Piperazine-2,5-diones as TGF-beta inhibitors

ABSTRACT

The present disclosure is concerned with piperazine-2,5-diones that are capable of inhibiting TGF-β and methods of treating cancers such as, for example, multiple myeloma and a hematologic malignancy, and methods of treating fibrotic conditions using these compounds. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

CROSS-REFERENCE TO RELATED APPLICATION

This Application is a national stage filing under 35 U.S.C. § 371 ofInternational Application No. PCT/US2019/045123, filed on Aug. 5, 2019,which claims the benefit of U.S. Application No. 62/715,733, filed onAug. 7, 2018, the contents of which are incorporated herein by referencein their entireties.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

This invention was made with government support under grant number1R01CA175012-01A awarded by the National Institutes of Health. Thegovernment has certain rights in the invention.

BACKGROUND

TGF-β is a central regulator of chronic liver disease through inductionof fibrogenic responses (Weiskirchen and Tacke (2016) Dig Dis34:410-422; Katz et al. (2016) Cancer letters 379:166-172; Yoshida etal. (2014) Int J Oncol 45:1363-1371; Fabregat et al. (2016) The FEBSjournal 283:2219-2232; Xu et al. (2016) J Histochem Cytochem64:157-167). Although infiltrating macrophages are a source of TGF-β,hepatic stellate cells are a significant source of TGF-β in liverfibrosis. TGF-β stimulates induction of myofibroblast-like properties ofhepatic stellate cells to produce extracellular matrix, leading tofibrosis. Although TGF-β inhibits hepatocyte proliferation under basalconditions, it has pro-oncogenic properties during malignant progressionthrough stimulating epithelial to mesenchymal transition, cell survivaland migration, and reduced immune surveillance.

TSP1 expression is increased in human liver disease with the THBS1 geneidentified as part of the characteristic gene signature of chronic liverdisease, including cirrhosis, in humans (Smalling et al. (2013) Am JPhysiol Gastrointest Liver Physiol 305:G364-374). In vitro studies showthat bile acids increase expression of TSP1 by hepatocytes, resulting inincreased TGF-β signaling in co-cultured hepatic stellate cells (Myunget al. (2007) Biochem Biophys Res Commun 353:1091-1096). Both TSP1 andTGF-β are increased in congenital hepatic fibrosis (El-Youssef (1999)Journal of pediatric gastroenterology and nutrition 28:386-392). TSP1regulated TGF-β activation prevented hepatocyte proliferation and liverregeneration after partial hepatectomy in Thbs1 deficient mice (Hayashiet al. (2012) Hepatology 55:1562-1573) and TSP1 induction by obstructedportal flow in mice is thought to lead to TGF-β-dependent liver atrophy(Hayashi et al. (2016) Hepatol Res 55: 1562-1573). TSP1 has been shownto regulate latent TGF-β activation in animal models of liver fibrosisand in cell culture models (reviewed in Li et al. (2016) Hepatol Res,doi: 10.1111/hepr.1287). Treatment of rats with the TSP1 antagonistpeptide LSKL prevented TGF-β activation and reduced liver fibrosis inthe dimethylnitrosamine model (Kondou et al. (2003) J Hepatol39:742-748). TSP1 is required for TGF-β signaling in both culturedhepatocytes and hepatic stellate cells, which is blocked by LSKL peptide(Breitkopf et al. (2005) Gut 54:673-681; Narmada (2013) J Cell Physiol228:393-401). Interestingly, TSP1-dependent latent TGF-β activationmight play a role in hepatitis C induced fibrosis and carcinogenesis asthe hepatitis C core protein induces TSP1 expression by hepatocytes toincrease active TGF-β and LSKL peptide blocks hepatitis C core proteinactivation of TGF-β (Benzoubir et al. (2013) J Hepatol 59:1160-1168).LSKL peptide administered early after injury also accelerated liverregeneration in mice following partial hepatectomy through blockingTGF-β activation and signaling (Kuroki et al. (2015) Br J Surg102:813-825). Both the TGF-β1 and the TGF-β2 isoforms are upregulated inmouse models and in human tissues with liver fibrosis and alsohepatocellular carcinoma (Dropmann et al. (2016) Oncotarget7:19499-19518): this is interesting since TSP1 can activate both the β1and β2 isoforms of latent TGF-β, whereas β2 cannot be activated byintegrin-dependent mechanisms.

Genetic ablation of TGF-β, its receptors, or its signaling mediatorsresults in developmental defects, inflammation, and increasedcarcinomas. Thus, it is therapeutically advantageous to target onlyadverse TGF-β activity in liver disease and spare homeostatic activity.Current anti-TGF-β therapeutics target the molecule itself or downstreamsignaling pathways and provide no mechanism for distinguishing betweenhomeostatic and disease-related TGF-β activity, thereby increasing thepotential for adverse effects. In fact, Smad 2 resistance and increasedpapilloma incidence in mice treated for 20 weeks with a TGF-β receptorkinase inhibitor have been identified (Connolly et al. (2011) Cancer Res71:2339-2349) and the 1D11 pan-specific anti-TGF-β neutralizing antibodyshows epithelial hyperplasia and progression to carcinoma in some models(Prud'homme (2007) Lab Invest 87:1077-1091).

TGF-β is secreted as a biologically inactive growth factor and controlof the conversion of latent TGF-β to a biologically active growth factoris a major regulatory node. Binding of the N-terminal latency associatedpeptide (LAP) prevents TGF-β binding to its receptors and thisinteraction must be disrupted for TGF-β signaling to occur. Latent TGF-βcan be converted to the active form through multiple mechanisms thatinclude proteolysis, binding to integrins, mechanical forces,modifications of the latent complex by viral enzymes or by reactiveoxygen species, or by binding to the secreted and ECM protein TSP1(Sweetwyne and Murphy-Ullrich (2012)Matrix Biol 31:178-186;Murphy-Ullrich and Poczatek (2000) Cytokine Growth Factor Rev 11:59-69).The mechanism that regulates latent TGF-β activation can vary withtissue, cell type, and specific disease milieu. Blockade of the majoractivation mechanism in a particular disease typically attenuatesadverse effects of TGF-β. Thus, it is important to identify thepredominant mechanism of TGF-β activation in multiple myeloma.

Thrombospondin 1 (TSP1) is a complex multi-functional protein releasedfrom platelet α-granules, incorporated into the fibrin clot, andexpressed by cell types that participate in wound healing responses in atemporally regulated manner (Agah et al. (2002) Am J Pathol 161:831-839;Murphy-Ullrich and Mosher (1985) Blood 66:1098-1104; DiPietro et al.(1996) Am J Pathol 148:1851-1860; Reed et al. (1993) J HistochemCytochem 41:1467-1477; Raugi et al. (1987) J Invest Dermatol89:551-554). TSP1 regulates multiple cellular events involved in tissuerepair including hemostasis, cell adhesion, migration, proliferation,ECM expression and organization, and regulation of growth factoractivity (Adams and Lawler (2004) Int J Biochem Cell Biol 36:961-968;Adams and Lawler (2011) Cold Spring Harb Perspect Biol 3:a009712). Inaddition to physiologic repair, TSP1 is also expressed at elevatedlevels in many tissues undergoing fibro-proliferative remodeling andblockade of specific actions of TSP1 or loss of TSP1 expression canattenuate pathologic tissue remodeling (Hugo (2003) Nephrol DialTransplant 18:1241-1245; Poczatek et al. (2000) Am J Pathol157:1353-1363; Daniel et al. (2007) Diabetes 56:2982-2989). TSP1 is amajor regulator of latent TGF-β activation (Murphy-Ullrich and Poczatek(2000) Cytokine Growth Factor Rev 11:59-69). TSP1 also hasTGF-β-independent functions in hemostasis, cell adhesion, migration, andgrowth factor regulation, e.g. regulation of epidermal growth factor(EGF), VEGF, and fibroblast growth factor (FGF) (Adams and Lawler (2011)Cold Spring Harb Perspect Biol 3:a009712). TSP1 is an endogenousangiogenesis inhibitor via inhibition of VEGF and FGF signaling. TSP1binding to Cluster of Differentiation 47 (CD47) and Cluster ofDifferentiation 36 (CD36) blocks nitric oxide signaling.

TSP1 is a secreted ECM protein that controls TGF-β activity by bindingand activating latent TGF-β (Sweetwyne and Murphy-Ullrich (2012) MatrixBiol 31:178-186; Murphy-Ullrich and Poczatek (2000) Cytokine GrowthFactor Rev 11:59-69). TSP1 binds to latent TGF-β to activate TGF-β atthe cell surface or in the extracellular milieu (Sweetwyne andMurphy-Ullrich (2012) Matrix Biol 31:178-186). Activation occurs throughbinding of the KRFK (-lysine-arginine-phenylalanine-lysine-) sequence inthe TSP1 type 1 repeats (TSRs) to LSKL (-leucine-serine-lysine-leucine-)in the LAP of the latent complex, which disrupts LAP-mature domaininteractions to expose the receptor binding sequences on the maturedomain, rendering TGF-β capable of signaling (Young and Murphy-Ullrich(2004) J Biol Chem 279:38032-38039). Peptide mimetics of sequencesinvolved in TSP1-TGF-β binding competitively inhibit TSP1-TGF-βactivation and studies with these peptides have established TSP1 as aprimary regulator of TGF-β bioactivity in different diseases (Sweetwyneand Murphy-Ullrich (2012)Matrix Biol 31:178-186). The tetrapeptide LSKL,which competitively blocks TSP-LAP binding, has been used in rodentmodels to inhibit TSP1-TGF-β activation and attenuate disease. Dosedependent intraperitoneal injection (i.p.) of LSKL improves end organfunction in murine diabetic nephropathy and rat cardiomyopathy byblocking TGF-β signaling in target tissues (Belmadani et al. (2007) Am JPathol 171:777-789; Lu et al. (2011) Am J Pathol 178:2573-2586). Animalsnecropsied after 15 weeks of treatment with 30 mg/kg i.p. LSKL, 3 timesweekly, showed no inflammation, no tumors in all major organs, and noimpairment of wound healing (Lu et al. (2011) Am J Pathol178:2573-2586).

In vitro studies have shown that TSP1 activates latent TGF-β secreted bymultiple cell types including endothelial cells, mesangial cells,hepatic stellate cells and skin, lung, and cardiac fibroblasts, T cells,and macrophages (Breitkopf et al. (2005) Gut 54:673-681; Murphy-Ullrichand Poczatek (2000) Cytokine Growth Factor Rev 11:59-69; et al. (2000)Am J Pathol 157:1353-1363; Mimura et al. (2005) Am J Pathol166:1451-1463; Yehualaeshet et al. (1999) Am J Pathol 155:841-851; Zhouet al. (2006) Biochem Biophys Res Commun 339:633-641; Schultz-Cherry andMurphy-Ullrich (1993) J Cell Biol 122:923-932; Yevdokimova et al. (2001)J Am Soc Nephrol 12:703-712; Yang et al. (2009) J Autoimmun 32: 94-103;Zhou et al. (2004) Am J Pathol 165:659-669). Peptides such as LSKL orWxxW which block TSP1 binding to the latent complex or antibodies whichblock TSP1-dependent TGF-β activation such as monoclonal antibody 133(Mab 133) have been used to establish the involvement of endogenous TSP1in TGF-β activation in a number of disease conditions and physiologicprocesses (Belmadani et al. (2007) Am J Pathol 171:777-789; Lu et al.(2011) Am J Pathol 178:2573-2586; Crawford et al. (1998) Cell93:1159-1170; Daniel et al. (2004) Kidney Int 65:459-468; Kondou et al.(2003) J Hepatol 39:742-748).

Initial evidence for an in vivo role of TSP1 in latent TGF-β activationwas shown by the ability of the KRFK peptide administered in theperinatal period to partially rescue the abnormal TSP-1 null phenotype,in particular airway epithelial hyperplasia and pancreatic islethyperplasia/acinar hypoplasia (Crawford et al. (1998) Cell93:1159-1170). Furthermore, treatment of wild type mice with the LSKLblocking peptide in the perinatal period replicated features of the TSP1knockout phenotype in the airways and pancreas. Double knockout of bothβ₆ integrin and TSP1 results in a phenotype distinct from either singleknockout that is characterized by severe inflammation, cardiacdegeneration, and epithelial hyperplasia, suggesting both separate andsynergistic roles in regulating latent TGF-β activation (Ludlow et al.(2005) J Cell Mol Med 9:421-437). However, it is likely that the primaryrole for TSP1 in controlling TGF-β activation is during injury, understress, and in pathologic conditions, rather than during development.The expression of TSP1 is induced by factors associated with systemicdiseases with fibrotic end organ involvement including high glucose,reactive oxygen species, and angiotensin II (Zhou et al. (2006) BiochemBiophys Res Commun 339:633-641; Yevdokimova et al. (2001) J Am SocNephrol 12:703-712; Wang et al. (2002) J Biol Chem 277:9880-9888; Wanget al. (2004) J Biol Chem 279:34311-34322). Indeed there is evidencefrom studies utilizing TSP1 antagonist peptides and diabetic TSP1knockout mice that TSP1 is a major factor in the development of fibroticend organ complications in diabetes (Daniel et al. (2007) Diabetes56:2982-2989; Belmadani et al. (2007) Am J Pathol 171:777-789; Lu et al.(2011) Am J Pathol 178:2573-2586). Treatment with i.p. injections ofLSKL, but not LSAL (leucine-serine-alanine-leucine) control peptide,reduced cardiac fibrosis, Smad phosphorylation, and improved leftventricular function (Belmadani et al. (2007) Am J Pathol 171:777-789).Similarly, treatment of Akita mice, a model of type 1 diabetes, withi.p. LSKL reduced urinary TGF-β activity and renal phospho-Smad 2/3levels and improved markers of tubulointerstitial injury and podocytefunction. (Lu et al. (2011) Am J Pathol 178:2573-2586). Interestingly,several studies have shown that TSP1 is involved in alveolarmacrophage-dependent TGF-β activation in mouse and rat models ofbleomycin-induced pulmonary fibrosis and treatment with either TSP1 orCD36 antagonist peptides can ameliorate lung fibrosis and reduce activeTGF-β (Chen et al. (2009) Exp. Toxicol. Pathol. 61: 59-65; Yehualaeshetet al. (2000) Am. J. Respir. Cell Mol. Biol. 23: 204-12).

One of the roles of TSP1 in dermal wound healing appears to beregulating the activation of latent TGF-β. The phenotype of excisionalwound healing in the TSP1 null mouse is consistent with a decrease inlocal TGF-β activation (Agah et al. (2002) Am J Pathol 161:831-839) andis characterized by a delay in macrophage recruitment and capillaryangiogenesis and a persistence of granulation tissue,neovascularization, and inflammation (Nor et al. (2005) Oral Biosci Med2:153-161). Topical treatment of TSP1 null wounds with the KRFKactivating peptide largely rescued the TSP1 null wound phenotype (Nor etal. (2005) Oral Biosci Med 2:153-161). TGF-β levels in these wounds wereincreased following KRFK treatment and the effects of the KRFK peptidewere blocked by a pan-specific anti-TGF-β antibody. While these datasuggest that TSP1 plays a role in local activation of TGF-β duringwounding, the studies of Agah et al., concluded that the decreasedactive and total TGF-β in the wounds of TSP1 or TSP1/TSP2 null mice isindirect and primarily due to defects in macrophage recruitment towounds (a major source of TGF-β in wounds) leading to an overallreduction in TGF-β rather than a defect in activation (Agah et al.(2002) Am J Pathol 161:831-839). Despite this controversy, it is clearthat TSP1 has the potential to modify the wound healing process.Subcutaneous implantation of TSP1 soaked sponges increased levels ofactive TGF-β, gel contraction and fibroblast migration (Sakai et al.(2003) J Dermatol Sci 31:99-109). Overexpression of TSP1 in keloids andin scleroderma correlates with increased TGF-β activity (Mimura et al.(2005) Am J Pathol 166:1451-1463; et al. (2000) Cell Death Differ7:166-176; Chen et al. (2011) Fibrogenesis Tissue Repair 4:9). Othershave used a derivative of the KRFK sequence, KFK(lysine-phenylalanine-lysine) coupled to a fatty acyl moiety to locallyactivate TGF-β and increase TIMP-1, which reduces MMP-induced elastinand collagen degradation when applied to dermal fibroblast cultures(Cauchard et al. (2004) Biochem Pharmacol 67:2013-2022). Systemicadministration of the LSKL blocking peptide did not reduce Smadsignaling or impair dermal wound healing in diabetic mice, although,these studies did not address the effects of direct LSKL administrationto the wounds and it is not known if local dermal levels of LSKLfollowing systemic intraperitoneal peptide administration are sufficientto alter local TGF-β activation (Lu et al. (2011) Am J Pathol178:2573-2586).

Although peptides comprising the amino acid sequence LSKL capable ofstimulating TGF-β activity are known, these peptides are often costlyand difficult to synthesize. Moreover, small molecules such as LSKL havean extremely short plasma stability half-life, only 2.1 minutes. Thus,there remains a need for small molecules capable of altering TGF-βactivity that are less expensive, easier to synthesize, and have anextended plasma stability half-life and methods of making and usingsame.

SUMMARY

In accordance with the purpose(s) of the invention, as embodied andbroadly described herein, the invention, in one aspect, relates topiperazine-2,5-diones compounds useful in the treatment of disordersassociated with a dysregulation of TGF-β including, but not limited to,cancers, immune dysfunction, and fibrotic conditions.

Disclosed are compounds having a structure represented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NR^(20a)R^(20b) and Cy¹; wherein each of R^(20a)and R^(20b), when present, is independently selected from hydrogen,C1-C4 alkyl, C1-C4 haloalkyl, cycloalkyl, and heteroaryl; wherein Cy¹,when present, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl,and —C₆H₄R²¹ and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; and wherein R²¹, when present, is selected from—(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b); wherein q, whenpresent, is selected from 0 and 1; wherein each of R^(31a), R^(31b),R^(32a), and R^(32b), when present, is independently selected fromhydrogen and C1-C4 alkyl; or wherein each of R^(1a) and R^(1b) ishydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), or wherein R^(1a) ishydrogen, R^(1b) is —CH₂Cy², and R⁴ is —C₆H₄R²¹; wherein Cy² is selectedfrom C3-C6 cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; provided that when n is 1 and each of R^(1a) and R^(1b)together comprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof.

Also disclosed are pharmaceutical compositions comprising atherapeutically effective amount of at least one disclosed compound anda pharmaceutically acceptable carrier.

Also disclosed are methods for inhibiting TGF-β activity in a subject,the method comprising the step of administering to the subject aneffective amount of at least one compound having a structure representedby a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); orwherein R^(1a) is hydrogen, R^(1b) is —CH₂Cy³ and R⁴ is —C₆H₄R²¹;wherein Cy³ is selected from C3-C6 cycloalkyl, monocyclic heteroaryl,and aryl and is substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; or a pharmaceutically acceptable salt thereof, therebyinhibiting TGF-β activity in the subject.

Also disclosed are methods for inhibiting TGF-β activity in a subject,the method comprising the step of administering to the subject aneffective amount of at least one compound having a structure selectedfrom:

or a pharmaceutically acceptable salt thereof, thereby inhibiting TGF-βactivity in the subject.

Also disclosed are methods for inhibiting TGF-β activity in at least onecell, the method comprising the step of contacting the cell with aneffective amount of at least one compound having a structure representedby a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), orwherein R^(1a) is hydrogen, R^(1b) is —CH₂Cy³ and R⁴ is —C₆H₄R²¹;wherein Cy³ is selected from C3-C6 cycloalkyl, monocyclic heteroaryl,and aryl and is substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; or a pharmaceutically acceptable salt thereof, therebyTGF-β activity in the cell.

Also disclosed are methods for inhibiting TGF-β activity in at least onecell, the method comprising the step of contacting the cell with aneffective amount of at least one compound selected from:

or a pharmaceutically acceptable salt thereof, thereby TGF-β activity inthe cell.

Also disclosed are kits comprising at least one disclosed compound andone or more of: (a) at least one agent known to increase TGF-β activity;(b) at least one agent known to treat cancer; (c) at least one agentknown to treat a fibrotic disorder; (d) at least one agent known totreat an immune dysfunction; (e) instructions for treating a disorderassociated with TGF-β dysfunction; (f) instructions for treating cancer;(g) instructions for treating a fibrotic disorder; and (h) instructionsfor treating an immune dysfunction.

Still other objects and advantages of the present disclosure will becomereadily apparent by those skilled in the art from the following detaileddescription, wherein it is shown and described only the preferredembodiments, simply by way of illustration of the best mode. As will berealized, the disclosure is capable of other and different embodiments,and its several details are capable of modifications in various obviousrespects, without departing from the disclosure. Accordingly, thedescription is to be regarded as illustrative in nature and not asrestrictive.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying figures, which are incorporated in and constitute apart of this specification, illustrate several aspects and together withthe description serve to explain the principles of the invention.

FIG. 1A and FIG. 1B show representative data illustrating the impact ofTSP1, LSKL and TGF-β on osteoblast differentiation by MSCs underosteogenic conditions.

FIG. 2A and FIG. 2B show representative data illustrating the impact ofLSKL treatment on tumor burden in the SCID-tibia MM model.

FIG. 3A-C show representative data illustrating the impact of LSKLtreatment on Smad 2 phosphorylation in bone marrow myeloma cells.

FIG. 4A and FIG. 4B show representative data illustrating the impact ofTSP1 and LSKL on TGF-β activity in CAG MM cells.

FIG. 5 shows a representative image of potentially significantinteractions in the binding pocket.

Additional advantages of the invention will be set forth in part in thedescription which follows, and in part will be obvious from thedescription, or can be learned by practice of the invention. Theadvantages of the invention will be realized and attained by means ofthe elements and combinations particularly pointed out in the appendedclaims. It is to be understood that both the foregoing generaldescription and the following detailed description are exemplary andexplanatory only and are not restrictive of the invention, as claimed.

DETAILED DESCRIPTION

The present invention can be understood more readily by reference to thefollowing detailed description of the invention and the Examplesincluded therein.

Before the present compounds, compositions, articles, systems, devices,and/or methods are disclosed and described, it is to be understood thatthey are not limited to specific synthetic methods unless otherwisespecified, or to particular reagents unless otherwise specified, as suchmay, of course, vary. It is also to be understood that the terminologyused herein is for the purpose of describing particular aspects only andis not intended to be limiting. Although any methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, example methods andmaterials are now described.

While aspects of the present invention can be described and claimed in aparticular statutory class, such as the system statutory class, this isfor convenience only and one of skill in the art will understand thateach aspect of the present invention can be described and claimed in anystatutory class. Unless otherwise expressly stated, it is in no wayintended that any method or aspect set forth herein be construed asrequiring that its steps be performed in a specific order. Accordingly,where a method claim does not specifically state in the claims ordescriptions that the steps are to be limited to a specific order, it isno way intended that an order be inferred, in any respect. This holdsfor any possible non-express basis for interpretation, including mattersof logic with respect to arrangement of steps or operational flow, plainmeaning derived from grammatical organization or punctuation, or thenumber or type of aspects described in the specification.

Throughout this application, various publications are referenced. Thedisclosures of these publications in their entireties are herebyincorporated by reference into this application in order to more fullydescribe the state of the art to which this pertains. The referencesdisclosed are also individually and specifically incorporated byreference herein for the material contained in them that is discussed inthe sentence in which the reference is relied upon. Nothing herein is tobe construed as an admission that the present invention is not entitledto antedate such publication by virtue of prior invention. Further, thedates of publication provided herein may be different from the actualpublication dates, which can require independent confirmation.

A. Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification, unless otherwise limited in specificinstances, either individually or as part of a larger group.

As used in the specification and the appended claims, the singular forms“a,” “an” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “a functionalgroup,” “an alkyl,” or “a residue” includes mixtures of two or more suchfunctional groups, alkyls, or residues, and the like.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another aspect includes from the one particular value and/orto the other particular value. Similarly, when values are expressed asapproximations, by use of the antecedent “about,” it will be understoodthat the particular value forms another aspect. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint. It is also understood that there are a number of valuesdisclosed herein, and that each value is also herein disclosed as“about” that particular value in addition to the value itself. Forexample, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that each unit between two particularunits are also disclosed. For example, if 10 and 15 are disclosed, then11, 12, 13, and 14 are also disclosed.

References in the specification and concluding claims to parts by weightof a particular element or component in a composition denotes the weightrelationship between the element or component and any other elements orcomponents in the composition or article for which a part by weight isexpressed. Thus, in a compound containing 2 parts by weight of componentX and 5 parts by weight component Y, X and Y are present at a weightratio of 2:5, and are present in such ratio regardless of whetheradditional components are contained in the compound.

A weight percent (wt. %) of a component, unless specifically stated tothe contrary, is based on the total weight of the formulation orcomposition in which the component is included.

As used herein, the terms “optional” or “optionally” mean that thesubsequently described event or circumstance can or cannot occur, andthat the description includes instances where said event or circumstanceoccurs and instances where it does not.

As used herein, the term “subject” can be a vertebrate, such as amammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject ofthe herein disclosed methods can be a human, non-human primate, horse,pig, rabbit, dog, sheep, goat, cow, cat, guinea pig, or rodent. The termdoes not denote a particular age or sex. Thus, adult and newbornsubjects, as well as fetuses, whether male or female, are intended to becovered. In one aspect, the subject is a mammal. A patient refers to asubject afflicted with a viral infection. The term “patient” includeshuman and veterinary subjects. In some aspects of the disclosed methods,the subject has been diagnosed with a need for treatment of one or moreviral infections prior to the administering step. In various aspects,the one or more disorders is selected from chikungunya, Venezuelanequine encephalitis, dengue, influenza, and zika.

As used herein, the term “treatment” refers to the medical management ofa patient with the intent to cure, ameliorate, stabilize, or prevent adisease, pathological condition, or disorder. This term includes activetreatment, that is, treatment directed specifically toward theimprovement of a disease, pathological condition, or disorder, and alsoincludes causal treatment, that is, treatment directed toward removal ofthe cause of the associated disease, pathological condition, ordisorder. In addition, this term includes palliative treatment, that is,treatment designed for the relief of symptoms rather than the curing ofthe disease, pathological condition, or disorder; preventativetreatment, that is, treatment directed to minimizing or partially orcompletely inhibiting the development of the associated disease,pathological condition, or disorder; and supportive treatment, that is,treatment employed to supplement another specific therapy directedtoward the improvement of the associated disease, pathologicalcondition, or disorder. In various aspects, the term covers anytreatment of a subject, including a mammal (e.g., a human), andincludes: (i) preventing the disease from occurring in a subject thatcan be predisposed to the disease but has not yet been diagnosed ashaving it; (ii) inhibiting the disease, i.e., arresting its development;or (iii) relieving the disease, i.e., causing regression of the disease.In one aspect, the subject is a mammal such as a primate, and, in afurther aspect, the subject is a human. The term “subject” also includesdomesticated animals (e.g., cats, dogs, etc.), livestock (e.g., cattle,horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mouse,rabbit, rat, guinea pig, fruit fly, etc.).

As used herein, the term “prevent” or “preventing” refers to precluding,averting, obviating, forestalling, stopping, or hindering something fromhappening, especially by advance action. It is understood that wherereduce, inhibit, or prevent are used herein, unless specificallyindicated otherwise, the use of the other two words is also expresslydisclosed.

As used herein, the term “diagnosed” means having been subjected to aphysical examination by a person of skill, for example, a physician, andfound to have a condition that can be diagnosed or treated by thecompounds, compositions, or methods disclosed herein. In some aspects ofthe disclosed methods, the subject has been diagnosed with a need fortreatment of a viral infection prior to the administering step. As usedherein, the phrase “identified to be in need of treatment for adisorder,” or the like, refers to selection of a subject based upon needfor treatment of the disorder. It is contemplated that theidentification can, in one aspect, be performed by a person differentfrom the person making the diagnosis. It is also contemplated, in afurther aspect, that the administration can be performed by one whosubsequently performed the administration.

As used herein, the terms “administering” and “administration” refer toany method of providing a pharmaceutical preparation to a subject. Suchmethods are well known to those skilled in the art and include, but arenot limited to, oral administration, transdermal administration,administration by inhalation, nasal administration, topicaladministration, intravaginal administration, ophthalmic administration,intraaural administration, intracerebral administration, rectaladministration, and parenteral administration, including injectable suchas intravenous administration, intra-arterial administration,intramuscular administration, and subcutaneous administration.Administration can be continuous or intermittent. In various aspects, apreparation can be administered therapeutically; that is, administeredto treat an existing disease or condition. In further various aspects, apreparation can be administered prophylactically; that is, administeredfor prevention of a disease or condition.

The term “treating” refers to relieving the disease, disorder, orcondition, i.e., causing regression of the disease, disorder, and/orcondition. The term “preventing” refers to preventing a disease,disorder, or condition from occurring in a human or an animal that maybe predisposed to the disease, disorder and/or condition, but has notyet been diagnosed as having it; and/or inhibiting the disease,disorder, or condition, i.e., arresting its development.

The term “contacting” as used herein refers to bringing a disclosedcompound and a cell, target receptor, or other biological entitytogether in such a manner that the compound can affect the activity ofthe target (e.g., receptor, cell, etc.), either directly; i.e., byinteracting with the target itself, or indirectly; i.e., by interactingwith another molecule, co-factor, factor, or protein on which theactivity of the target is dependent.

As used herein, the terms “effective amount” and “amount effective”refer to an amount that is sufficient to achieve the desired result orto have an effect on an undesired condition. For example, a“therapeutically effective amount” refers to an amount that issufficient to achieve the desired therapeutic result or to have aneffect on undesired symptoms, but is generally insufficient to causeadverse side effects. The specific therapeutically effective dose levelfor any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the specific composition employed; the age, body weight, general health,sex and diet of the patient; the time of administration; the route ofadministration; the rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed and like factors well known in themedical arts. For example, it is well within the skill of the art tostart doses of a compound at levels lower than those required to achievethe desired therapeutic effect and to gradually increase the dosageuntil the desired effect is achieved. If desired, the effective dailydose can be divided into multiple doses for purposes of administration.Consequently, single dose compositions can contain such amounts orsubmultiples thereof to make up the daily dose. The dosage can beadjusted by the individual physician in the event of anycontraindications. Dosage can vary, and can be administered in one ormore dose administrations daily, for one or several days. Guidance canbe found in the literature for appropriate dosages for given classes ofpharmaceutical products. In further various aspects, a preparation canbe administered in a “prophylactically effective amount”; that is, anamount effective for prevention of a disease or condition.

As used herein, “IC₅₀,” is intended to refer to the concentration of asubstance (e.g., a compound or a drug) that is required for 50%inhibition of a biological process, or component of a process, includinga protein, subunit, organelle, ribonucleoprotein, etc. In one aspect, anIC₅₀ can refer to the concentration of a substance that is required for50% inhibition in vivo, as further defined elsewhere herein.

The term “comprising” (and its grammatical variations) as used herein isused in the inclusive sense of “having” or “including” and not in theexclusive sense of “consisting of.”

The compounds according to this disclosure may form prodrugs at hydroxylor amino functionalities using alkoxy, amino acids, etc., groups as theprodrug forming moieties. For instance, the hydroxymethyl position mayform mono-, di- or triphosphates and again these phosphates can formprodrugs. Preparations of such prodrug derivatives are discussed invarious literature sources (examples are: Alexander et al., J. Med.Chem. 1988, 31, 318; Aligas-Martin et al., PCT WO 2000/041531, p. 30).The nitrogen function converted in preparing these derivatives is one(or more) of the nitrogen atoms of a compound of the disclosure.

“Derivatives” of the compounds disclosed herein are pharmaceuticallyacceptable salts, prodrugs, deuterated forms, radio-actively labeledforms, isomers, solvates and combinations thereof. The “combinations”mentioned in this context are refer to derivatives falling within atleast two of the groups: pharmaceutically acceptable salts, prodrugs,deuterated forms, radio-actively labeled forms, isomers, and solvates.Examples of radio-actively labeled forms include compounds labeled withtritium, phosphorous-32, iodine-129, carbon-11, fluorine-18, and thelike.

“Pharmaceutically acceptable salts” refer to derivatives of thedisclosed compounds wherein the parent compound is modified by makingacid or base salts thereof. The compounds of this disclosure form acidaddition salts with a wide variety of organic and inorganic acids andinclude the physiologically acceptable salts which are often used inpharmaceutical chemistry. Such salts are also part of this disclosure.Typical inorganic acids used to form such salts include hydrochloric,hydrobromic, hydroiodic, nitric, sulfuric, phosphoric, hypophosphoricacid, and the like. Salts derived from organic acids, such as aliphaticmono- and dicarboxylic acids, phenyl substituted alkanoic acids,hydroxyalkanoic and hydroxyalkandioic acids, aromatic acids, aliphaticand aromatic sulfonic acids may also be used. Such pharmaceuticallyacceptable salts thus include acetate, phenylacetate, trifluoroacetate,acrylate, ascorbate, benzoate, chlorobenzoate, dinitrobenzoate,hydroxybenzoate, methoxybenzoate, methylbenzoate, o-acetoxybenzoate,naphthalene-2-benzoate, bromide, isobutyrate, phenylbutyrate,β-hydroxybutyrate, butyne-1,4-dioate, hexyne-1,4-dioate, caprate,caprylate, chloride, cinnamate, citrate, formate, fumarate, glycollate,heptanoate, hippurate, lactate, malate, maleate, hydroxymaleate,malonate, mandelate, mesylate, nicotinate, isonicotinate, nitrate,oxalate, phthalate, teraphthalate, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, propiolate,propionate, phenylpropionate, salicylate, sebacate, succinate, suberate,sulfate, bisulfate, pyrosulfate, sulfite, bisulfite, sulfonate,benzene-sulfonate, p-bromobenzenesulfonate, chlorobenzenesulfonate,ethanesulfonate, 2-hydroxyethanesulfonate, methanesulfonate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, p-toleunesulfonate,xylenesulfonate, tartarate, and the like.

It is understood that the compounds of the present disclosure relate toall optical isomers and stereo-isomers at the various possible atoms ofthe molecule, unless specified otherwise. Compounds may be separated orprepared as their pure enantiomers or diasteriomers by crystallization,chromatography or synthesis.

The term “leaving group” refers to an atom (or a group of atoms) withelectron withdrawing ability that can be displaced as a stable species,taking with it the bonding electrons. Examples of suitable leavinggroups include sulfonate esters, including triflate, mesylate, tosylate,brosylate, and halides.

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, and aromatic and nonaromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described below. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of this disclosure, the heteroatoms, such as nitrogen, canhave hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valences of theheteroatoms. This disclosure is not intended to be limited in any mannerby the permissible substituents of organic compounds. Also, the terms“substitution” or “substituted with” include the implicit proviso thatsuch substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., a compound that does not spontaneouslyundergo transformation such as by rearrangement, cyclization,elimination, etc. It is also contemplated that, in certain aspects,unless expressly indicated to the contrary, individual substituents canbe further optionally substituted (i.e., further substituted orunsubstituted).

In defining various terms, “A¹,” “A²,” “A³,” and “A⁴” are used herein asgeneric symbols to represent various specific substituents. Thesesymbols can be any substituent, not limited to those disclosed herein,and when they are defined to be certain substituents in one instance,they can, in another instance, be defined as some other substituents.

The term “alkyl” as used herein is a branched or unbranched saturatedhydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl,n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl,isopentyl, s-pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl,dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, and the like. Thealkyl group can also be substituted or unsubstituted. The alkyl groupcan be substituted with one or more groups including, but not limitedto, optionally substituted alkyl, cycloalkyl, alkoxy, amino, ether,halide, hydroxy, nitro, silyl, sulfo-oxo, or thiol, as described herein.A “lower alkyl” group is an alkyl group containing from one to six(e.g., from one to four) carbon atoms.

Throughout the specification “alkyl” is generally used to refer to bothunsubstituted alkyl groups and substituted alkyl groups; however,substituted alkyl groups are also specifically referred to herein byidentifying the specific substituent(s) on the alkyl group. For example,the term “halogenated alkyl” specifically refers to an alkyl group thatis substituted with one or more halide, e.g., fluorine, chlorine,bromine, or iodine. The term “alkoxyalkyl” specifically refers to analkyl group that is substituted with one or more alkoxy groups, asdescribed below. The term “alkylamino” specifically refers to an alkylgroup that is substituted with one or more amino groups, as describedbelow, and the like. When “alkyl” is used in one instance and a specificterm such as “alkylalcohol” is used in another, it is not meant to implythat the term “alkyl” does not also refer to specific terms such as“alkylalcohol” and the like.

This practice is also used for other groups described herein. That is,while a term such as “cycloalkyl” refers to both unsubstituted andsubstituted cycloalkyl moieties, the substituted moieties can, inaddition, be specifically identified herein; for example, a particularsubstituted cycloalkyl can be referred to as, e.g., an“alkylcycloalkyl.” Similarly, a substituted alkoxy can be specificallyreferred to as, e.g., a “halogenated alkoxy,” a particular substitutedalkenyl can be, e.g., an “alkenylalcohol,” and the like. Again, thepractice of using a general term, such as “cycloalkyl,” and a specificterm, such as “alkylcycloalkyl,” is not meant to imply that the generalterm does not also include the specific term.

The term “cycloalkyl” as used herein is a non-aromatic carbon-based ringcomposed of at least three carbon atoms. Examples of cycloalkyl groupsinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, norbornyl, and the like. The term “heterocycloalkyl” is atype of cycloalkyl group as defined above, and is included within themeaning of the term “cycloalkyl,” where at least one of the carbon atomsof the ring is replaced with a heteroatom such as, but not limited to,nitrogen, oxygen, sulfur, or phosphorus. The cycloalkyl group andheterocycloalkyl group can be substituted or unsubstituted. Thecycloalkyl group and heterocycloalkyl group can be substituted with oneor more groups including, but not limited to, optionally substitutedalkyl, cycloalkyl, alkoxy, amino, ether, halide, hydroxy, nitro, silyl,sulfo-oxo, or thiol as described herein.

The term “polyalkylene group” as used herein is a group having two ormore CH₂ groups linked to one another. The polyalkylene group can berepresented by the formula —(CH₂)_(a)—, where “a” is an integer of from2 to 500.

The terms “alkoxy” and “alkoxyl” as used herein to refer to an alkyl orcycloalkyl group bonded through an ether linkage; that is, an “alkoxy”group can be defined as —OA¹ where A¹ is alkyl or cycloalkyl as definedabove. “Alkoxy” also includes polymers of alkoxy groups as justdescribed; that is, an alkoxy can be a polyether such as —OA¹-OA² or—OA¹-(OA²)_(a)-OA³, where “a” is an integer of from 1 to 200 and A¹, A²,and A³ are alkyl and/or cycloalkyl groups.

The term “alkenyl” as used herein is a hydrocarbon group of from 2 to 24carbon atoms with a structural formula containing at least onecarbon-carbon double bond. Asymmetric structures such as (A¹A²)C═C(A³A⁴)are intended to include both the E and Z isomers. This can be presumedin structural formulae herein wherein an asymmetric alkene is present,or it can be explicitly indicated by the bond symbol C═C. The alkenylgroup can be substituted with one or more groups including, but notlimited to, optionally substituted alkyl, cycloalkyl, alkoxy, alkenyl,cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino,carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro,silyl, sulfo-oxo, or thiol, as described herein.

The term “cycloalkenyl” as used herein is a non-aromatic carbon-basedring composed of at least three carbon atoms and containing at least onecarbon-carbon double bound, i.e., C═C. Examples of cycloalkenyl groupsinclude, but are not limited to, cyclopropenyl, cyclobutenyl,cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl,norbornenyl, and the like. The term “heterocycloalkenyl” is a type ofcycloalkenyl group as defined above, and is included within the meaningof the term “cycloalkenyl,” where at least one of the carbon atoms ofthe ring is replaced with a heteroatom such as, but not limited to,nitrogen, oxygen, sulfur, or phosphorus. The cycloalkenyl group andheterocycloalkenyl group can be substituted or unsubstituted. Thecycloalkenyl group and heterocycloalkenyl group can be substituted withone or more groups including, but not limited to, optionally substitutedalkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl,aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether,halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol asdescribed herein.

The term “alkynyl” as used herein is a hydrocarbon group of 2 to 24carbon atoms with a structural formula containing at least onecarbon-carbon triple bond. The alkynyl group can be unsubstituted orsubstituted with one or more groups including, but not limited to,optionally substituted alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl,alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino, carboxylicacid, ester, ether, halide, hydroxy, ketone, azide, nitro, silyl,sulfo-oxo, or thiol, as described herein.

The term “cycloalkynyl” as used herein is a non-aromatic carbon-basedring composed of at least seven carbon atoms and containing at least onecarbon-carbon triple bound. Examples of cycloalkynyl groups include, butare not limited to, cycloheptynyl, cyclooctynyl, cyclononynyl, and thelike. The term “heterocycloalkynyl” is a type of cycloalkenyl group asdefined above, and is included within the meaning of the term“cycloalkynyl,” where at least one of the carbon atoms of the ring isreplaced with a heteroatom such as, but not limited to, nitrogen,oxygen, sulfur, or phosphorus. The cycloalkynyl group andheterocycloalkynyl group can be substituted or unsubstituted. Thecycloalkynyl group and heterocycloalkynyl group can be substituted withone or more groups including, but not limited to, optionally substitutedalkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl,aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether,halide, hydroxy, ketone, azide, nitro, silyl, sulfo-oxo, or thiol asdescribed herein.

The term “aryl” as used herein is a group that contains any carbon-basedaromatic group including, but not limited to, benzene, naphthalene,phenyl, biphenyl, phenoxybenzene, and the like. The term “aryl” alsoincludes “heteroaryl,” which is defined as a group that contains anaromatic group that has at least one heteroatom incorporated within thering of the aromatic group. Examples of heteroatoms include, but are notlimited to, nitrogen, oxygen, sulfur, and phosphorus. Likewise, the term“non-heteroaryl,” which is also included in the term “aryl,” defines agroup that contains an aromatic group that does not contain aheteroatom. The aryl group can be substituted or unsubstituted. The arylgroup can be substituted with one or more groups including, but notlimited to, optionally substituted alkyl, cycloalkyl, alkoxy, alkenyl,cycloalkenyl, alkynyl, cycloalkynyl, aryl, heteroaryl, aldehyde, amino,carboxylic acid, ester, ether, halide, hydroxy, ketone, azide, nitro,silyl, sulfo-oxo, or thiol as described herein. The term “biaryl” is aspecific type of aryl group and is included in the definition of “aryl.”Biaryl refers to two aryl groups that are bound together via a fusedring structure, as in naphthalene, or are attached via one or morecarbon-carbon bonds, as in biphenyl.

The term “aldehyde” as used herein is represented by the formula —C(O)H.Throughout this specification “C(O)” is a short hand notation for acarbonyl group, i.e., C═O.

The terms “amine” or “amino” as used herein are represented by theformula —NA¹A², where A¹ and A² can be, independently, hydrogen oralkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl,or heteroaryl group as described herein.

The term “alkylamino” as used herein is represented by the formula—NH(-alkyl) where alkyl is a described herein. Representative examplesinclude, but are not limited to, methylamino group, ethylamino group,propylamino group, isopropylamino group, butylamino group, isobutylaminogroup, (sec-butyl)amino group, (tert-butyl)amino group, pentylaminogroup, isopentylamino group, (tert-pentyl)amino group, hexylamino group,and the like.

The term “dialkylamino” as used herein is represented by the formula—N(-alkyl)₂ where alkyl is a described herein. Representative examplesinclude, but are not limited to, dimethylamino group, diethylaminogroup, dipropylamino group, diisopropylamino group, dibutylamino group,diisobutylamino group, di(sec-butyl)amino group, di(tert-butyl)aminogroup, dipentylamino group, diisopentylamino group, di(tert-pentyl)aminogroup, dihexylamino group, N-ethyl-N-methylamino group,N-methyl-N-propylamino group, N-ethyl-N-propylamino group and the like.

The term “carboxylic acid” as used herein is represented by the formula—C(O)OH.

The term “ester” as used herein is represented by the formula —OC(O)A¹or —C(O)OA¹, where A¹ can be an optionally substituted alkyl,cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, orheteroaryl group as described herein. The term “polyester” as usedherein is represented by the formula -(A¹O(O)C-A²-C(O)O)_(a)— or-(A¹O(O)C-A²-OC(O))_(a)—, where A¹ and A² can be, independently, anoptionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, cycloalkynyl, aryl, or heteroaryl group described herein and“a” is an integer from 1 to 500. “Polyester” is as the term used todescribe a group that is produced by the reaction between a compoundhaving at least two carboxylic acid groups with a compound having atleast two hydroxyl groups.

The term “ether” as used herein is represented by the formula A¹OA²,where A¹ and A² can be, independently, an optionally substituted alkyl,cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, orheteroaryl group described herein. The term “polyether” as used hereinis represented by the formula -(A¹O-A²O)_(a)—, where A¹ and A² can be,independently, an optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, cycloalkynyl, aryl, or heteroaryl group describedherein and “a” is an integer of from 1 to 500. Examples of polyethergroups include polyethylene oxide, polypropylene oxide, and polybutyleneoxide.

The term “halide” as used herein refers to the halogens fluorine,chlorine, bromine, and iodine.

The term “heterocycle,” as used herein refers to single and multi-cyclicaromatic or non-aromatic ring systems in which at least one of the ringmembers is other than carbon. Heterocycle includes pyridine, pyrimidine,furan, thiophene, pyrrole, isoxazole, isothiazole, pyrazole, oxazole,thiazole, imidazole, oxazole, including, 1,2,3-oxadiazole,1,2,5-oxadiazole and 1,3,4-oxadiazole, thiadiazole, including,1,2,3-thiadiazole, 1,2,5-thiadiazole, and 1,3,4-thiadiazole, triazole,including, 1,2,3-triazole, 1,3,4-triazole, tetrazole, including1,2,3,4-tetrazole and 1,2,4,5-tetrazole, pyridine, pyridazine,pyrimidine, pyrazine, triazine, including 1,2,4-triazine and1,3,5-triazine, tetrazine, including 1,2,4,5-tetrazine, pyrrolidine,piperidine, piperazine, morpholine, azetidine, tetrahydropyran,tetrahydrofuran, dioxane, and the like.

The term “hydroxyl” as used herein is represented by the formula —OH.

The term “ketone” as used herein is represented by the formula A¹C(O)A²,where A¹ and A² can be, independently, an optionally substituted alkyl,cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, orheteroaryl group as described herein.

The term “azide” as used herein is represented by the formula —N₃.

The term “nitro” as used herein is represented by the formula —NO₂.

The term “nitrile” as used herein is represented by the formula —CN.

The term “silyl” as used herein is represented by the formula -SiA¹A²A³,where A¹, A², and A³ can be, independently, hydrogen or an optionallysubstituted alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, alkynyl,cycloalkynyl, aryl, or heteroaryl group as described herein.

The term “sulfo-oxo” as used herein is represented by the formulas—S(O)A¹, —S(O)₂A¹, —OS(O)₂A¹, or —OS(O)₂OA¹, where A¹ can be hydrogen oran optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.Throughout this specification “S(O)” is a short hand notation for S═O.The term “sulfonyl” is used herein to refer to the sulfo-oxo grouprepresented by the formula —S(O)₂A¹, where A¹ can be hydrogen or anoptionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl,alkynyl, cycloalkynyl, aryl, or heteroaryl group as described herein.The term “sulfone” as used herein is represented by the formulaA¹S(O)₂A², where A¹ and A² can be, independently, an optionallysubstituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl,cycloalkynyl, aryl, or heteroaryl group as described herein. The term“sulfoxide” as used herein is represented by the formula A¹S(O)A², whereA¹ and A² can be, independently, an optionally substituted alkyl,cycloalkyl, alkenyl, cycloalkenyl, alkynyl, cycloalkynyl, aryl, orheteroaryl group as described herein.

The term “thiol” as used herein is represented by the formula —SH.

“R¹,” “R²,” “R³,” “R,” where n is an integer, as used herein can,independently, possess one or more of the groups listed above. Forexample, if R¹ is a straight chain alkyl group, one of the hydrogenatoms of the alkyl group can optionally be substituted with a hydroxylgroup, an alkoxy group, an alkyl group, a halide, and the like.Depending upon the groups that are selected, a first group can beincorporated within second group or, alternatively, the first group canbe pendant (i.e., attached) to the second group. For example, with thephrase “an alkyl group comprising an amino group,” the amino group canbe incorporated within the backbone of the alkyl group. Alternatively,the amino group can be attached to the backbone of the alkyl group. Thenature of the group(s) that is (are) selected will determine if thefirst group is embedded or attached to the second group.

As described herein, compounds of the invention may contain “optionallysubstituted” moieties. In general, the term “substituted,” whetherpreceded by the term “optionally” or not, means that one or morehydrogens of the designated moiety are replaced with a suitablesubstituent. Unless otherwise indicated, an “optionally substituted”group may have a suitable substituent at each substitutable position ofthe group, and when more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds. In is also contemplated that, in certain aspects,unless expressly indicated to the contrary, individual substituents canbe further optionally substituted (i.e., further substituted orunsubstituted).

The term “stable,” as used herein, refers to compounds that are notsubstantially altered when subjected to conditions to allow for theirproduction, detection, and, in certain aspects, their recovery,purification, and use for one or more of the purposes disclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group are independently halogen;—(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O(CH₂)₀₋₄R^(∘), —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘);—(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted with R^(∘); —NO₂;—CN; —N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘);—N(R^(∘))C(S)R^(∘); —(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘)₂; —(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃; —(CH₂)₀₋₄OC(O)R^(∘);—OC(O)(CH₂)₀₋₄SR—, SC(S)SR^(∘); —(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR^(∘)₂; —C(S)NR^(∘) ₂; —C(S)SR^(∘); —SC(S)SR^(∘), —(CH₂)₀₋₄OC(O)NR^(∘) ₂;—C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘);—C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘); —(CH₂)₀₋₄S(O)₂R^(∘);—(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘); —S(O)₂NR^(∘) ₂;—(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂; —N(R^(∘))S(O)₂R^(∘);—N(OR^(∘))R^(∘); —C(NH)NR^(∘) ₂; —P(O)₂R^(∘); —P(O)R^(∘) ₂; —OP(O)R^(∘)₂; —OP(O)(OR^(∘))₂; SiR^(∘) ₃; —(C1-4 straight or branchedalkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight or branchedalkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substituted asdefined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5-6 membered heteroaryl ring), or a 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(∘), taken together with their intervening atom(s), form a3-12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R^(∘) (or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), are independently halogen, —(CH₂)O₂R^(●), -(haloR^(●)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(●), —(CH₂)₀₋₂CH(OR^(●))₂; —O(haloR^(●)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(●), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(●),—(CH₂)₀₋₂SR^(●), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(●),—(CH₂)₀₋₂NR^(●) ₂, —NO₂, —SiR^(●)3, —OSiR^(●) ₃, —C(O)SR^(●), —(C₁₋₄straight or branched alkylene)C(O)OR^(●), or —SSR^(●) wherein each R^(●)is unsubstituted or where preceded by “halo” is substituted only withone or more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O, ═S, ═NNR*₂,═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or—S(C(R*₂))₂₋₃S—, wherein each independent occurrence of R* is selectedfrom hydrogen, C₁₋₆ aliphatic which may be substituted as defined below,or an unsubstituted 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. Suitable divalent substituents that are bound tovicinal substitutable carbons of an “optionally substituted” groupinclude: —O(CR*₂)₂₋₃O—, wherein each independent occurrence of R* isselected from hydrogen, C₁₋₆ aliphatic which may be substituted asdefined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN, —C(O)OH,—C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein each R^(●) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein eachR^(†) is independently hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R^(†) are independentlyhalogen, —R^(●), -(haloR^(●)), —OH, —OR^(●), —O(haloR^(●)), —CN,—C(O)OH, —C(O)OR^(●), —NH₂, —NHR^(●), —NR^(●) ₂, or —NO₂, wherein eachR^(●) is unsubstituted or where preceded by “halo” is substituted onlywith one or more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

The term “organic residue” defines a carbon containing residue, i.e., aresidue comprising at least one carbon atom, and includes but is notlimited to the carbon-containing groups, residues, or radicals definedhereinabove. Organic residues can contain various heteroatoms, or bebonded to another molecule through a heteroatom, including oxygen,nitrogen, sulfur, phosphorus, or the like. Examples of organic residuesinclude but are not limited alkyl or substituted alkyls, alkoxy orsubstituted alkoxy, mono or di-substituted amino, amide groups, etc.Organic residues can preferably comprise 1 to 18 carbon atoms, 1 to 15,carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbonatoms, or 1 to 4 carbon atoms. In a further aspect, an organic residuecan comprise 2 to 18 carbon atoms, 2 to 15, carbon atoms, 2 to 12 carbonatoms, 2 to 8 carbon atoms, 2 to 4 carbon atoms, or 2 to 4 carbon atoms

A very close synonym of the term “residue” is the term “radical,” whichas used in the specification and concluding claims, refers to afragment, group, or substructure of a molecule described herein,regardless of how the molecule is prepared. For example, a2,4-thiazolidinedione radical in a particular compound has thestructure:

regardless of whether thiazolidinedione is used to prepare the compound.In some embodiments the radical (for example an alkyl) can be furthermodified (i.e., substituted alkyl) by having bonded thereto one or more“substituent radicals.” The number of atoms in a given radical is notcritical to the present invention unless it is indicated to the contraryelsewhere herein.

“Organic radicals,” as the term is defined and used herein, contain oneor more carbon atoms. An organic radical can have, for example, 1-26carbon atoms, 1-18 carbon atoms, 1-12 carbon atoms, 1-8 carbon atoms,1-6 carbon atoms, or 1-4 carbon atoms. In a further aspect, an organicradical can have 2-26 carbon atoms, 2-18 carbon atoms, 2-12 carbonatoms, 2-8 carbon atoms, 2-6 carbon atoms, or 2-4 carbon atoms. Organicradicals often have hydrogen bound to at least some of the carbon atomsof the organic radical. One example, of an organic radical thatcomprises no inorganic atoms is a 5, 6, 7, 8-tetrahydro-2-naphthylradical. In some embodiments, an organic radical can contain 1-10inorganic heteroatoms bound thereto or therein, including halogens,oxygen, sulfur, nitrogen, phosphorus, and the like. Examples of organicradicals include but are not limited to an alkyl, substituted alkyl,cycloalkyl, substituted cycloalkyl, mono-substituted amino,di-substituted amino, acyloxy, cyano, carboxy, carboalkoxy,alkylcarboxamide, substituted alkylcarboxamide, dialkylcarboxamide,substituted dialkylcarboxamide, alkylsulfonyl, alkylsulfinyl, thioalkyl,thiohaloalkyl, alkoxy, substituted alkoxy, haloalkyl, haloalkoxy, aryl,substituted aryl, heteroaryl, heterocyclic, or substituted heterocyclicradicals, wherein the terms are defined elsewhere herein. A fewnon-limiting examples of organic radicals that include heteroatomsinclude alkoxy radicals, trifluoromethoxy radicals, acetoxy radicals,dimethylamino radicals and the like.

“Inorganic radicals,” as the term is defined and used herein, contain nocarbon atoms and therefore comprise only atoms other than carbon.Inorganic radicals comprise bonded combinations of atoms selected fromhydrogen, nitrogen, oxygen, silicon, phosphorus, sulfur, selenium, andhalogens such as fluorine, chlorine, bromine, and iodine, which can bepresent individually or bonded together in their chemically stablecombinations. Inorganic radicals have 10 or fewer, or preferably one tosix or one to four inorganic atoms as listed above bonded together.Examples of inorganic radicals include, but not limited to, amino,hydroxy, halogens, nitro, thiol, sulfate, phosphate, and like commonlyknown inorganic radicals. The inorganic radicals do not have bondedtherein the metallic elements of the periodic table (such as the alkalimetals, alkaline earth metals, transition metals, lanthanide metals, oractinide metals), although such metal ions can sometimes serve as apharmaceutically acceptable cation for anionic inorganic radicals suchas a sulfate, phosphate, or like anionic inorganic radical. Inorganicradicals do not comprise metalloids elements such as boron, aluminum,gallium, germanium, arsenic, tin, lead, or tellurium, or the noble gaselements, unless otherwise specifically indicated elsewhere herein.

Compounds described herein can contain one or more double bonds and,thus, potentially give rise to cis/trans (E/Z) isomers, as well as otherconformational isomers. Unless stated to the contrary, the inventionincludes all such possible isomers, as well as mixtures of such isomers.

Unless stated to the contrary, a formula with chemical bonds shown onlyas solid lines and not as wedges or dashed lines contemplates eachpossible isomer, e.g., each enantiomer and diastereomer, and a mixtureof isomers, such as a racemic or scalemic mixture. Compounds describedherein can contain one or more asymmetric centers and, thus, potentiallygive rise to diastereomers and optical isomers. Unless stated to thecontrary, the present invention includes all such possible diastereomersas well as their racemic mixtures, their substantially pure resolvedenantiomers, all possible geometric isomers, and pharmaceuticallyacceptable salts thereof. Mixtures of stereoisomers, as well as isolatedspecific stereoisomers, are also included. During the course of thesynthetic procedures used to prepare such compounds, or in usingracemization or epimerization procedures known to those skilled in theart, the products of such procedures can be a mixture of stereoisomers.

Many organic compounds exist in optically active forms having theability to rotate the plane of plane-polarized light. In describing anoptically active compound, the prefixes D and L or R and S are used todenote the absolute configuration of the molecule about its chiralcenter(s). The prefixes d and l or (+) and (−) are employed to designatethe sign of rotation of plane-polarized light by the compound, with (−)or meaning that the compound is levorotatory. A compound prefixed with(+) or d is dextrorotatory. For a given chemical structure, thesecompounds, called stereoisomers, are identical except that they arenon-superimposable mirror images of one another. A specific stereoisomercan also be referred to as an enantiomer, and a mixture of such isomersis often called an enantiomeric mixture. A 50:50 mixture of enantiomersis referred to as a racemic mixture. Many of the compounds describedherein can have one or more chiral centers and therefore can exist indifferent enantiomeric forms. If desired, a chiral carbon can bedesignated with an asterisk (*). When bonds to the chiral carbon aredepicted as straight lines in the disclosed formulas, it is understoodthat both the (R) and (S) configurations of the chiral carbon, and henceboth enantiomers and mixtures thereof, are embraced within the formula.As is used in the art, when it is desired to specify the absoluteconfiguration about a chiral carbon, one of the bonds to the chiralcarbon can be depicted as a wedge (bonds to atoms above the plane) andthe other can be depicted as a series or wedge of short parallel linesis (bonds to atoms below the plane). The Cahn-Inglod-Prelog system canbe used to assign the (R) or (S) configuration to a chiral carbon.

When the disclosed compounds contain one chiral center, the compoundsexist in two enantiomeric forms. Unless specifically stated to thecontrary, a disclosed compound includes both enantiomers and mixtures ofenantiomers, such as the specific 50:50 mixture referred to as a racemicmixture. The enantiomers can be resolved by methods known to thoseskilled in the art, such as formation of diastereoisomeric salts whichmay be separated, for example, by crystallization (see, CRC Handbook ofOptical Resolutions via Diastereomeric Salt Formation by David Kozma(CRC Press, 2001)); formation of diastereoisomeric derivatives orcomplexes which may be separated, for example, by crystallization,gas-liquid or liquid chromatography; selective reaction of oneenantiomer with an enantiomer-specific reagent, for example enzymaticesterification; or gas-liquid or liquid chromatography in a chiralenvironment, for example on a chiral support for example silica with abound chiral ligand or in the presence of a chiral solvent. It will beappreciated that where the desired enantiomer is converted into anotherchemical entity by one of the separation procedures described above, afurther step can liberate the desired enantiomeric form. Alternatively,specific enantiomers can be synthesized by asymmetric synthesis usingoptically active reagents, substrates, catalysts or solvents, or byconverting one enantiomer into the other by asymmetric transformation.

Designation of a specific absolute configuration at a chiral carbon in adisclosed compound is understood to mean that the designatedenantiomeric form of the compounds can be provided in enantiomericexcess (e.e.). Enantiomeric excess, as used herein, is the presence of aparticular enantiomer at greater than 50%, for example, greater than60%, greater than 70%, greater than 75%, greater than 80%, greater than85%, greater than 90%, greater than 95%, greater than 98%, or greaterthan 99%. In one aspect, the designated enantiomer is substantially freefrom the other enantiomer. For example, the “R” forms of the compoundscan be substantially free from the “S” forms of the compounds and are,thus, in enantiomeric excess of the “S” forms. Conversely, “S” forms ofthe compounds can be substantially free of “R” forms of the compoundsand are, thus, in enantiomeric excess of the “R” forms.

When a disclosed compound has two or more chiral carbons, it can havemore than two optical isomers and can exist in diastereoisomeric forms.For example, when there are two chiral carbons, the compound can have upto four optical isomers and two pairs of enantiomers ((S,S)/(R,R) and(R,S)/(S,R)). The pairs of enantiomers (e.g., (S,S)/(R,R)) are mirrorimage stereoisomers of one another. The stereoisomers that are notmirror-images (e.g., (S,S) and (R,S)) are diastereomers. Thediastereoisomeric pairs can be separated by methods known to thoseskilled in the art, for example chromatography or crystallization andthe individual enantiomers within each pair may be separated asdescribed above. Unless otherwise specifically excluded, a disclosedcompound includes each diastereoisomer of such compounds and mixturesthereof.

Compounds described herein comprise atoms in both their natural isotopicabundance and in non-natural abundance. The disclosed compounds can beisotopically-labeled or isotopically-substituted compounds identical tothose described, but for the fact that one or more atoms are replaced byan atom having an atomic mass or mass number different from the atomicmass or mass number typically found in nature. Examples of isotopes thatcan be incorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine,such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ¹⁸F and ³⁶Cl,respectively. Compounds further comprise prodrugs thereof, andpharmaceutically acceptable salts of said compounds or of said prodrugswhich contain the aforementioned isotopes and/or other isotopes of otheratoms are within the scope of this invention. Certainisotopically-labeled compounds of the present invention, for examplethose into which radioactive isotopes such as ³H and ¹⁴C areincorporated, are useful in drug and/or substrate tissue distributionassays. Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes areparticularly preferred for their ease of preparation and detectability.Further, substitution with heavier isotopes such as deuterium, i.e., ²H,can afford certain therapeutic advantages resulting from greatermetabolic stability, for example increased in vivo half-life or reduceddosage requirements and, hence, may be preferred in some circumstances.Isotopically labeled compounds of the present invention and prodrugsthereof can generally be prepared by carrying out the procedures below,by substituting a readily available isotopically labeled reagent for anon-isotopically labeled reagent.

The compounds described in the invention can be present as a solvate.“Solvates” refers to the compound formed by the interaction of a solventand a solute and includes hydrates. Solvates are usually crystallinesolid adducts containing solvent molecules within the crystal structure,in either stoichiometric or nonstoichiometric proportions. In somecases, the solvent used to prepare the solvate is an aqueous solution,and the solvate is then often referred to as a hydrate. The compoundscan be present as a hydrate, which can be obtained, for example, bycrystallization from a solvent or from aqueous solution. In thisconnection, one, two, three or any arbitrary number of solvate or watermolecules can combine with the compounds according to the invention toform solvates and hydrates. Unless stated to the contrary, the inventionincludes all such possible solvates.

The term “co-crystal” means a physical association of two or moremolecules which owe their stability through non-covalent interaction.One or more components of this molecular complex provide a stableframework in the crystalline lattice. In certain instances, the guestmolecules are incorporated in the crystalline lattice as anhydrates orsolvates, see e.g. “Crystal Engineering of the Composition ofPharmaceutical Phases. Do Pharmaceutical Co-crystals Represent a NewPath to Improved Medicines?” Almarasson, O., et. al., The Royal Societyof Chemistry, 1889-1896, 2004. Examples of co-crystals includep-toluenesulfonic acid and benzenesulfonic acid.

It is known that chemical substances form solids which are present indifferent states of order which are termed polymorphic forms ormodifications. The different modifications of a polymorphic substancecan differ greatly in their physical properties. The compounds accordingto the invention can be present in different polymorphic forms, with itbeing possible for particular modifications to be metastable. Unlessstated to the contrary, the invention includes all such possiblepolymorphic forms.

In some aspects, a structure of a compound can be represented by aformula:

which is understood to be equivalent to a formula:

wherein n is typically an integer. That is, R^(n) is understood torepresent five independent substituents, R^(n(a)), R^(n(b)), R^(n(e)),R^(n(d)), R^(n(e)). In each such case, each of the five R^(n) can behydrogen or a recited substituent. By “independent substituents,” it ismeant that each R substituent can be independently defined. For example,if in one instance R^(n(a)) is halogen, then R^(n(b)) is not necessarilyhalogen in that instance.

In some yet further aspects, a structure of a compound can berepresented by a formula:

wherein R^(y) represents, for example, 0-2 independent substituentsselected from A¹, A², and A³, which is understood to be equivalent tothe groups of formulae:

-   -   wherein R^(y) represents 0 independent substituents

-   -   wherein R^(y) represents 1 independent substituent

-   -   wherein R^(y) represents 2 independent substituents

Again, by “independent substituents,” it is meant that each Rsubstituent can be independently defined. For example, if in oneinstance R^(y1) is A¹, then R^(y2) is not necessarily A¹ in thatinstance.

In some further aspects, a structure of a compound can be represented bya formula,

wherein, for example, Q comprises three substituents independentlyselected from hydrogen and A, which is understood to be equivalent to aformula:

Again, by “independent substituents,” it is meant that each Qsubstituent is independently defined as hydrogen or A, which isunderstood to be equivalent to the groups of formulae:

-   -   wherein Q comprises three substituents independently selected        from H and A

Certain materials, compounds, compositions, and components disclosedherein can be obtained commercially or readily synthesized usingtechniques generally known to those of skill in the art. For example,the starting materials and reagents used in preparing the disclosedcompounds and compositions are either available from commercialsuppliers such as Aldrich Chemical Co., (Milwaukee, Wis.), AcrosOrganics (Morris Plains, N.J.), Fisher Scientific (Pittsburgh, Pa.), orSigma (St. Louis, Mo.) or are prepared by methods known to those skilledin the art following procedures set forth in references such as Fieserand Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wileyand Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 andSupplementals (Elsevier Science Publishers, 1989); Organic Reactions,Volumes 1-40 (John Wiley and Sons, 1991); March's Advanced OrganicChemistry, (John Wiley and Sons, 4th Edition); and Larock'sComprehensive Organic Transformations (VCH Publishers Inc., 1989).

Unless otherwise expressly stated, it is in no way intended that anymethod set forth herein be construed as requiring that its steps beperformed in a specific order. Accordingly, where a method claim doesnot actually recite an order to be followed by its steps or it is nototherwise specifically stated in the claims or descriptions that thesteps are to be limited to a specific order, it is no way intended thatan order be inferred, in any respect. This holds for any possiblenon-express basis for interpretation, including: matters of logic withrespect to arrangement of steps or operational flow; plain meaningderived from grammatical organization or punctuation; and the number ortype of embodiments described in the specification.

Disclosed are the components to be used to prepare the compositions ofthe invention as well as the compositions themselves to be used withinthe methods disclosed herein. These and other materials are disclosedherein, and it is understood that when combinations, subsets,interactions, groups, etc. of these materials are disclosed that whilespecific reference of each various individual and collectivecombinations and permutation of these compounds cannot be explicitlydisclosed, each is specifically contemplated and described herein. Forexample, if a particular compound is disclosed and discussed and anumber of modifications that can be made to a number of moleculesincluding the compounds are discussed, specifically contemplated is eachand every combination and permutation of the compound and themodifications that are possible unless specifically indicated to thecontrary. Thus, if a class of molecules A, B, and C are disclosed aswell as a class of molecules D, E, and F and an example of a combinationmolecule, A-D is disclosed, then even if each is not individuallyrecited each is individually and collectively contemplated meaningcombinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considereddisclosed. Likewise, any subset or combination of these is alsodisclosed. Thus, for example, the sub-group of A-E, B-F, and C-E wouldbe considered disclosed. This concept applies to all aspects of thisapplication including, but not limited to, steps in methods of makingand using the compositions of the invention. Thus, if there are avariety of additional steps that can be performed it is understood thateach of these additional steps can be performed with any specificembodiment or combination of embodiments of the methods of theinvention.

It is understood that the compositions disclosed herein have certainfunctions. Disclosed herein are certain structural requirements forperforming the disclosed functions, and it is understood that there area variety of structures that can perform the same function that arerelated to the disclosed structures, and that these structures willtypically achieve the same result.

B. Compounds

In one aspect, disclosed are compounds useful in treating disordersassociated with dysregulation of TGF-β, in particular, cancers, immunedysfunction, and fibrotic conditions.

In one aspect, the disclosed compounds exhibit inhibition of TGF-β.

In one aspect, the disclosed compounds are useful in inhibiting TGF-β ina mammal. In a further aspect, the disclosed compounds are useful ininhibiting TGF-β activity in at least one cell.

In one aspect, the disclosed compounds are useful in the treatment ofcancers, as further described herein.

In one aspect, the disclosed compounds are useful in the treatment offibrotic conditions, as further described herein.

In one aspect, the disclosed compounds are useful in the treatment ofimmune dysfunction, as described herein.

It is contemplated that each disclosed derivative can be optionallyfurther substituted. It is also contemplated that any one or morederivative can be optionally omitted from the invention. It isunderstood that a disclosed compound can be provided by the disclosedmethods. It is also understood that the disclosed compounds can beemployed in the disclosed methods of using.

1. Structure

In one aspect, disclosed are compounds having a structure represented bya formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NR^(20a)R^(20b) and Cy¹; wherein each of R^(20a)and R^(20b), when present, is independently selected from hydrogen,C1-C4 alkyl, C1-C4 haloalkyl, cycloalkyl, and heteroaryl; wherein Cy¹,when present, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl,and —C₆H₄R²¹ and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; and wherein R²¹, when present, is selected from—(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b); wherein q, whenpresent, is selected from 0 and 1; wherein each of R^(31a), R^(31b),R^(32a), and R^(32b), when present, is independently selected fromhydrogen and C1-C4 alkyl; or wherein each of R^(1a) and R^(1b) ishydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), or wherein R^(1a) ishydrogen, R^(1b) is —CH₂Cy², and R⁴ is —C₆H₄R²¹; wherein Cy² is selectedfrom C3-C6 cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; provided that when n is 1 and each of R^(1a) and R^(1b)together comprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof.

In one aspect, disclosed are compounds having a structure represented bya formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NR^(20a)R^(20b) and Cy¹; wherein each of R^(20a)and R^(20b), when present, is independently selected from hydrogen,C1-C4 alkyl, C1-C4 haloalkyl, cycloalkyl, and heteroaryl; wherein Cy¹,when present, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl,and —C₆H₄R²¹ and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; and wherein R²¹, when present, is selected from—(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b); wherein q, whenpresent, is selected from 0 and 1; wherein each of R^(31a), R^(31b),R^(32a), and R^(32b), when present, is independently selected fromhydrogen and C1-C4 alkyl; or wherein each of R^(1a) and R^(1b) ishydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), provided that when n is 1and each of R^(1a) and R^(1b) together comprise a 3-membered cycloalkylthen Cy¹, when present, is not monocyclic heteroaryl, and provided thatwhen R⁴ is —NR^(20a)R^(20b) then each of R^(1a) and R^(1b) isindependently C1-C4 alkyl, or a pharmaceutically acceptable saltthereof.

In a further aspect, the compound has a structure represented by aformula selected from:

In a further aspect, the compound has a structure represented by aformula:

In a further aspect, the compound has a structure represented by aformula:

In a further aspect, the compound has a structure represented by aformula selected from:

In a further aspect, the compound has a structure represented by aformula:

In a further aspect, a structure represented by a formula:

wherein Z is selected from O, S, and NR⁴⁰; wherein R⁴⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein each of Q¹ and Q² isindependently selected from CH and N; and wherein R³³ is selected fromhydrogen and C1-C4 alkyl.

In a further aspect, the compound has a structure represented by aformula:

In a further aspect, the compound has a structure represented by aformula selected from:

In a further aspect, the compound has a structure represented by aformula:

wherein R³³ is selected from hydrogen and C1-C4 alkyl.

In a further aspect, the compound has a structure represented by aformula:

wherein Q¹ and Q² is independently selected from CH and N; and whereinR³³ is selected from hydrogen and C1-C4 alkyl.

In a further aspect, the compound has a structure selected from:

In a further aspect, the compound has a structure selected from:

In one aspect, n is selected from 1, 2, 3, and 4. In a further aspect, nis selected from 1, 2, and 3. In a still further aspect, n is selectedfrom 1 and 2. In yet a further aspect, n is 4. In an even furtheraspect, n is 3. In a still further aspect, n is 2. In yet a furtheraspect, n is 1.

In one aspect, q, when present, is selected from 0 and 1. In a furtheraspect, q, when present, is 0. In a still further aspect, q, whenpresent, is 1.

a. Q¹ and Q² Groups

In one aspect, each of Q¹ and Q² is independently selected from CH andN. In a further aspect, each of Q¹ and Q² is CH. In a still furtheraspect, each of Q¹ and Q² is N.

In a further aspect, Q¹ is CH and Q² is N. In a still further aspect, Q¹is N and Q² is CH.

b. Z Groups

In a further aspect, Z is selected from O, S, and NR⁴⁰. In a stillfurther aspect, Z is O. In yet a further aspect, Z is S. In an evenfurther aspect, Z is NR⁴⁰.

c. R^(1A) and R^(1B) Groups

In one aspect, each of R^(1a) and R^(1b) is independently C1-C4 alkyl;or each of R^(1a) and R^(1b) are optionally covalently bonded togetherand, together with the intermediate atoms, comprise a 3- to 7-memberedcycloalkyl substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; or each of R^(1a) and R^(1b) is hydrogen; or R^(1a) ishydrogen, R^(1b) is —CH₂Cy², and R⁴ is —C₆H₄R²¹.

In a further aspect, each of R^(1a) and R^(1b) is independently selectedfrom methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, andt-butyl. In a still further aspect, each of R^(1a) and R^(1b) isindependently selected from methyl, ethyl, n-propyl, and i-propyl. Inyet a further aspect, each of R^(1a) and R^(1b) is independentlyselected from methyl and ethyl. In an even further aspect, each ofR^(1a) and R^(1b) is ethyl. In a still further aspect, each of R^(1a)and R^(1b) is methyl.

In a further aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a 3-to 7-membered cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-3 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 3- to 7-memberedcycloalkyl substituted with 0-2 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-1 non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl,C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, each of R^(1a)and R^(1b) are optionally covalently bonded together and, together withthe intermediate atoms, comprise a 3- to 7-membered cycloalkylmonosubstituted with a non-hydrogen group selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise an unsubstituted 3- to7-membered cycloalkyl.

In a further aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a 3-to 6-membered cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 5-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 3- to 4-memberedcycloalkyl substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 4- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 5- to 7-memberedcycloalkyl substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 6- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 4- to 6-memberedcycloalkyl substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino.

In a further aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a4-membered cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 3-membered cycloalkyl substituted with0-3 non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 4-membered cycloalkylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a4-membered cycloalkyl substituted with 0-1 non-hydrogen group selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In astill further aspect, each of R^(1a) and R^(1b) are optionallycovalently bonded together and, together with the intermediate atoms,comprise a 4-membered cycloalkyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise an unsubstituted 4-membered cycloalkyl.

In a further aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a5-membered cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 5-membered cycloalkyl substituted with0-3 non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 5-membered cycloalkylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a5-membered cycloalkyl substituted with 0-1 non-hydrogen group selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In astill further aspect, each of R^(1a) and R^(1b) are optionallycovalently bonded together and, together with the intermediate atoms,comprise a 5-membered cycloalkyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise an unsubstituted 5-membered cycloalkyl.

In a further aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a6-membered cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise a 6-membered cycloalkyl substituted with0-3 non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,each of R^(1a) and R^(1b) are optionally covalently bonded together and,together with the intermediate atoms, comprise a 6-membered cycloalkylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, each of R^(1a) and R^(1b) are optionally covalentlybonded together and, together with the intermediate atoms, comprise a6-membered cycloalkyl substituted with 0-1 non-hydrogen group selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In astill further aspect, each of R^(1a) and R^(1b) are optionallycovalently bonded together and, together with the intermediate atoms,comprise a 6-membered cycloalkyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, each of R^(1a) and R^(1b) areoptionally covalently bonded together and, together with theintermediate atoms, comprise an unsubstituted 6-membered cycloalkyl.

In a further aspect, each of R^(1a) and R^(1b) is hydrogen.

In a further aspect, R^(1a) is hydrogen and R^(1b) is —CH₂Cy².

d. R² and R³ Groups

In one aspect, each of R² and R³ is independently selected from hydrogenand C1-C4 alkyl. In a further aspect, each of R² and R³ is hydrogen.

In a further aspect, each of R² and R³ is independently selected fromhydrogen, methyl, ethyl, n-propyl, and i-propyl. In a still furtheraspect, each of R² and R³ is independently selected from hydrogen,methyl, and ethyl. In yet a further aspect, each of R² and R³ isindependently selected from hydrogen and ethyl. In an even furtheraspect, each of R² and R³ is independently selected from hydrogen andmethyl.

In a further aspect, each of R² and R³ is independently C1-C4 alkyl. Ina still further aspect, each of R² and R³ is independently selected frommethyl, ethyl, n-propyl, and i-propyl. In yet a further aspect, each ofR² and R³ is independently selected from methyl and ethyl. In an evenfurther aspect, each of R² and R³ is ethyl. In a still further aspect,each of R² and R³ is methyl.

e. R⁴ Groups

In one aspect, R⁴ is selected from —NR^(20a)R^(20b) and Cy¹; or R⁴ is—C₆H₄C(O)NR^(32a)R^(32b).

In a further aspect, R⁴ is selected from —NR^(20a)R^(20b) and Cy¹. In astill further aspect, R⁴ is —NR^(20a)R^(20b). In yet a further aspect,R⁴ is —NHR^(20a). In an even further aspect, R⁴ is —NH₂. In an evenfurther aspect, R⁴ is Cy¹.

In a further aspect, R⁴ is —C₆H₄C(O)NR^(32a)R^(32b).

f. R^(20a) and R^(20b) Groups

In one aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, C1-C4 alkyl, C1-C4 haloalkyl,cycloalkyl, and heteroaryl. In a further aspect, each of R^(20a) andR^(20b), when present, is hydrogen.

In a further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, C1-C4 alkyl, and C1-C4 haloalkyl.In a still further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, methyl, ethyl, n-propyl, i-propyl,—CH₂Cl, —CH₂CH₂Cl, —CH₂CH₂CH₂Cl, —CH(CH₂C1)(CH₃), —CH₂F, —CH₂CH₂F,—CH₂CH₂CH₂F, and —CH(CH₂F)(CH₃). In yet a further aspect, each ofR^(20a) and R^(20b), when present, is independently selected fromhydrogen, methyl, ethyl, —CH₂Cl, —CH₂CH₂Cl, —CH₂F, and —CH₂CH₂F. In aneven further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, methyl, —CH₂Cl, and —CH₂F.

In a further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen and C1-C4 alkyl. In a still furtheraspect, each of R^(20a) and R^(20b), when present, is independentlyselected from hydrogen, methyl, ethyl, n-propyl, and i-propyl. In yet afurther aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, methyl, and ethyl. In an evenfurther aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen and ethyl. In a still furtheraspect, each of R^(20a) and R^(20b), when present, is independentlyselected from hydrogen and methyl.

In a further aspect, each of R^(20a) and R^(20b), when present, is C1-C4alkyl. In a still further aspect, each of R^(20a) and R^(20b), whenpresent, is independently selected from methyl, ethyl, n-propyl, andi-propyl. In yet a further aspect, each of R^(20a) and R^(20b), whenpresent, is independently selected from methyl and ethyl. In an evenfurther aspect, each of R^(20a) and R^(20b), when present, is ethyl. Ina still further aspect, each of R^(20a) and R^(20b), when present, ismethyl.

In a further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen and C1-C4 haloalkyl. In a stillfurther aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, —CH₂Cl, —CH₂CH₂Cl, —CH₂CH₂CH₂Cl,—CH(CH₂C1)(CH₃), —CH₂F, —CH₂CH₂F, —CH₂CH₂CH₂F, and —CH(CH₂F)(CH₃). Inyet a further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, —CH₂Cl, —CH₂CH₂Cl, —CH₂F, and—CH₂CH₂F. In an even further aspect, each of R^(20a) and R^(20b), whenpresent, is independently selected from hydrogen, —CH₂Cl, and —CH₂F.

In a further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, cycloalkyl, and heteroaryl. In astill further aspect, each of R^(20a) and R^(20b), when present, isindependently selected from hydrogen, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl,

g. R²¹ Groups

In one aspect, R²¹, when present, is selected from—(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b). In a further aspect,R²¹, when present, is —(CH₂)_(q)NR^(31a)R^(31b). In a still furtheraspect, R²¹, when present, is —C(O)NR^(32a)R^(32b).

h. R^(31a), R^(31b), R^(32a), and R^(32b) Groups

In one aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b), whenpresent, is independently selected from hydrogen and C1-C4 alkyl. In afurther aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b), whenpresent, is hydrogen.

In a further aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b),when present, is independently selected from hydrogen, methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a stillfurther aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b), whenpresent, is independently selected from hydrogen, methyl, ethyl,n-propyl, and i-propyl. In yet a further aspect, each of R^(31a),R^(31b), R^(32a), and R^(32b), when present, is independently selectedfrom hydrogen, methyl, and ethyl. In an even further aspect, each ofR^(31a), R^(31b), R^(32a) and R^(32b), when present, is independentlyselected from hydrogen and ethyl. In a still further aspect, each ofR^(31a), R^(31b), R^(32a), and R^(32b), when present, is independentlyselected from hydrogen and methyl.

In a further aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b),when present, is independently selected from methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a still furtheraspect, each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from methyl, ethyl, n-propyl, and i-propyl. Inyet a further aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b),when present, is independently selected from methyl, and ethyl. In aneven further aspect, each of R^(31a), R^(31b), R^(32a), and R^(32b),when present, is ethyl. In a still further aspect, each of R^(31a),R^(31b), R^(32a), and R^(32b), when present, is methyl.

i. R³³ Groups

In one aspect, R³³ is selected from hydrogen and C1-C4 alkyl. In afurther aspect, R³³ is hydrogen.

In a further aspect, R³³ is selected from hydrogen, methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a stillfurther aspect, R³³ is selected from hydrogen, methyl, ethyl, n-propyl,and i-propyl. In yet a further aspect, R³³ is selected from hydrogen,methyl, and ethyl. In an even further aspect, R³³ is selected fromhydrogen and ethyl. In a still further aspect, R³³ is selected fromhydrogen and methyl.

In a further aspect, R³³ is selected from methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a still furtheraspect, R³³ is selected from methyl, ethyl, n-propyl, and i-propyl. Inyet a further aspect, R³³ is selected from methyl and ethyl. In an evenfurther aspect, R³³ is ethyl. In a still further aspect, R³³ is methyl.

j. R⁴⁰ Groups

In one aspect, R⁴⁰, when present, is selected from hydrogen and C1-C4alkyl. In a further aspect, R⁴⁰ is hydrogen.

In a further aspect, R⁴⁰ is selected from hydrogen, methyl, ethyl,n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a stillfurther aspect, R⁴⁰ is selected from hydrogen, methyl, ethyl, n-propyl,and i-propyl. In yet a further aspect, R⁴⁰ is selected from hydrogen,methyl, and ethyl. In an even further aspect, R⁴⁰ is selected fromhydrogen and ethyl. In a still further aspect, R⁴⁰ is selected fromhydrogen and methyl.

In a further aspect, R⁴⁰ is selected from methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, s-butyl, and t-butyl. In a still furtheraspect, R⁴⁰ is selected from methyl, ethyl, n-propyl, and i-propyl. Inyet a further aspect, R⁴⁰ is selected from methyl and ethyl. In an evenfurther aspect, R⁴⁰ is ethyl. In a still further aspect, R⁴⁰ is methyl.

k. Cy¹ Groups

In one aspect, Cy¹, when present, is selected from C5-C6 cycloalkyl,monocyclic heteroaryl, and —C₆H₄R²¹ and is substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a further aspect, Cy¹,when present, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl,and —C₆H₄R²¹ and is substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, Cy¹, when present, is selectedfrom C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ and issubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy¹, when present, is selected from C5-C6 cycloalkyl,monocyclic heteroaryl, and —C₆H₄R²¹ and is substituted with 0-1non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In an even further aspect, Cy¹, whenpresent, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl, and—C₆H₄R²¹ and is monosubstituted with a non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy¹, when present, is selected from C5-C6 cycloalkyl,monocyclic heteroaryl, and —C₆H₄R²¹ and is unsubstituted.

In a further aspect, Cy¹, when present, is selected from C5-C6cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ and is monosubstitutedwith a group selected from —NH₂, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, Cy¹, when present, is selectedfrom C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ and ismonosubstituted with a group selected from —NH₂ and C1-C4 alkylamino. Inyet a further aspect, Cy¹, when present, is selected from C5-C6cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ and is monosubstitutedwith a —NH₂ group.

In a further aspect, Cy¹, when present, is C5-C6 cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy¹, when present, is C5-C6 cycloalkyl substituted with 0-3 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In yet a further aspect, Cy¹, when present,is C5-C6 cycloalkyl substituted with 0-2 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy¹, when present, is C5-C6cycloalkyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy¹, when present, is C5-C6 cycloalkyl monosubstitutedwith a non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl,C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In yet a further aspect, Cy¹, when present,is unsubstituted C5-C6 cycloalkyl.

In a further aspect, Cy¹, when present, is selected from cyclopentyl andcyclohexyl and is substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, Cy¹, when present, iscyclopentyl substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy¹, when present, is cyclohexylsubstituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino.

In a further aspect, Cy¹ is monocyclic heteroaryl substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy¹ is monocyclic heteroaryl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy¹ is monocyclic heteroarylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, Cy¹ is monocyclic heteroaryl substituted with 0-1non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy¹ ismonocyclic heteroaryl monosubstituted with a group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy¹ is unsubstituted monocyclic heteroaryl.

In a further aspect, Cy¹ is selected from pyridinyl, thiophenyl,furanyl, oxazolyl, thiazolyl, and 1,2,4-thiazolyl and is substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy¹ is selected from pyridinyl, oxazolyl, thiazolyl, and 1,2,4-thiazolyland is substituted with 0-4 non-hydrogen groups independently selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yeta further aspect, Cy¹ is selected from oxazolyl, thiazolyl, and1,2,4-thiazolyl and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy¹ is pyridinyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino.

In a further aspect, Cy¹ is —C₆H₄R²¹ substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy¹ is —C₆H₄R²¹substituted with 0-3 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy¹ is —C₆H₄R²¹ substituted with 0-2 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy¹ is —C₆H₄R²¹ substitutedwith 0-1 non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl,C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy¹ is —C₆H₄R²¹monosubstituted with a non-hydrogen group selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,Cy¹ is unsubstituted —C₆H₄R²¹.

In a further aspect, Cy¹ is —C₆H₄NH₂. In a still further aspect, Cy¹ is—C₆H₄CH₂NH₂.

In a further aspect, Cy¹ is monocyclic heteroaryl substituted with agroup selected from —(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b).In a still further aspect, Cy¹ is monocyclic heteroaryl substituted witha group selected from —NH₂ and —C(O)NH₂. In yet a further aspect, Cy¹ ismonocyclic heteroaryl substituted with a —C(O)NH₂ group. In an evenfurther aspect, Cy¹ is monocyclic heteroaryl substituted with a —NH₂group.

l. Cy² Groups

In one aspect, Cy² is selected from C3-C6 cycloalkyl substituted with0-4 non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a further aspect, Cy² isselected from C3-C6 cycloalkyl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, Cy² is selected from C3-C6cycloalkyl substituted with 0-2 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is selected from C3-C6cycloalkyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, Cy² is selected from C3-C6 cycloalkyl monosubstitutedwith a non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl,C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy² isunsubstituted C3-C6 cycloalkyl.

In a further aspect, Cy² is cyclobutyl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy² iscyclobutyl substituted with 0-3 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is cyclobutyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy² is cyclobutyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy² is cyclobutyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is unsubstituted cyclobutyl.

In a further aspect, Cy² is cyclopentyl substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy² is cyclopentyl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is cyclopentyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy² is cyclopentyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy² is cyclopentyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is unsubstituted cyclopentyl.

In a further aspect, Cy² is cyclohexyl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy² iscyclohexyl substituted with 0-3 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is cyclohexyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy² is cyclohexyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy² is cyclohexyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy² is unsubstituted cyclohexyl.

m. Cy³ Groups

In one aspect, Cy³ is selected from C3-C6 cycloalkyl, monocyclicheteroaryl, and aryl and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a further aspect, Cy³ is selected from C3-C6cycloalkyl, monocyclic heteroaryl, and aryl and is substituted with 0-3non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy³ is selected from C3-C6 cycloalkyl, monocyclic heteroaryl, and aryland is substituted with 0-2 non-hydrogen groups independently selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yeta further aspect, Cy³ is selected from C3-C6 cycloalkyl, monocyclicheteroaryl, and aryl and is substituted with 0-1 non-hydrogen groupselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy³ is selected from C3-C6cycloalkyl, monocyclic heteroaryl, and aryl and is monosubstituted witha non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is selectedfrom C3-C6 cycloalkyl, monocyclic heteroaryl, and aryl and isunsubstituted.

In a further aspect, Cy³ is C3-C6 cycloalkyl substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy³ is C3-C6 cycloalkyl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is C3-C6 cycloalkylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, Cy³ is C3-C6 cycloalkyl substituted with 0-1non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is C3-C6cycloalkyl monosubstituted with a non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy³ is unsubstituted C3-C6 cycloalkyl.

In a further aspect, Cy³ is cyclobutyl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ isCyclobutyl substituted with 0-3 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is cyclobutyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy³ is Cyclobutyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy³ is cyclobutyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is unsubstituted cyclobutyl.

In a further aspect, Cy³ is cyclopentyl substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy³ is Cyclopentyl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is cyclopentyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy³ is Cyclopentyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy³ is cyclopentyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is unsubstituted cyclopentyl.

In a further aspect, Cy³ is cyclohexyl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ isCyclohexyl substituted with 0-3 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is cyclohexyl substitutedwith 0-2 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an even further aspect,Cy³ is Cyclohexyl substituted with 0-1 non-hydrogen group selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a stillfurther aspect, Cy³ is cyclohexyl monosubstituted with a non-hydrogengroup selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is unsubstituted cyclohexyl.

In a further aspect, Cy³ is selected from monocyclic heteroaryl and aryland is substituted with 0-4 non-hydrogen groups independently selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In astill further aspect, Cy³ is selected from monocyclic heteroaryl andaryl and is substituted with 0-3 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is selected from monocyclicheteroaryl and aryl and is substituted with 0-2 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy³ is selected from monocyclicheteroaryl and aryl and is substituted with 0-1 non-hydrogen groupselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In a still further aspect, Cy³ is selected from monocyclicheteroaryl and aryl and is monosubstituted with a non-hydrogen groupselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is selected from monocyclicheteroaryl and aryl and is unsubstituted.

In a further aspect, Cy³ is monocyclic heteroaryl substituted with 0-4non-hydrogen groups independently selected from halogen, —NH₂, C1-C4alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In a still further aspect,Cy³ is Monocyclic heteroaryl substituted with 0-3 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In yet a further aspect, Cy³ is monocyclic heteroarylsubstituted with 0-2 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In an evenfurther aspect, Cy³ is Monocyclic heteroaryl substituted with 0-1non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ ismonocyclic heteroaryl monosubstituted with a non-hydrogen group selectedfrom halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl,C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yeta further aspect, Cy³ is unsubstituted monocyclic heteroaryl.

In a further aspect, Cy³ is aryl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is Arylsubstituted with 0-3 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy³ is aryl substituted with 0-2 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy³ is Aryl substituted with0-1 non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is arylmonosubstituted with a non-hydrogen group selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,Cy³ is unsubstituted aryl.

In a further aspect, Cy³ is C6-aryl substituted with 0-4 non-hydrogengroups independently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is C6-arylsubstituted with 0-3 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet afurther aspect, Cy³ is C6-aryl substituted with 0-2 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino. In an even further aspect, Cy³ is C6-aryl substituted with0-1 non-hydrogen group selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino. In a still further aspect, Cy³ is C6-arylmonosubstituted with a non-hydrogen group selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino. In yet a further aspect,Cy³ is unsubstituted C6-aryl.

2. Example Compounds

In one aspect, a compound can be present as one or more of the followingstructures:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be present as one or more of the followingstructures:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be present as one or more of the followingstructures:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be present as one or more of the followingstructures:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be present as:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound cannot be:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound cannot be:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound cannot be:

or a pharmaceutically acceptable salt thereof.

3. Prophetic Compound Examples

The following compound examples are prophetic, and can be prepared usingthe synthesis methods described herein above and other general methodsas needed as would be known to one skilled in the art. It is anticipatedthat the prophetic compounds would be active as inhibitors of TGF-β, andsuch activity can be determined using the assay methods describedherein.

In one aspect, a compound can be selected from:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be selected from:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be selected from:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be selected from:

or a pharmaceutically acceptable salt thereof.

In one aspect, a compound can be selected from:

or a pharmaceutically acceptable salt thereof.

C. Pharmaceutical Compositions

In one aspect, the invention relates to pharmaceutical compositionscomprising at least one disclosed compound and a pharmaceuticallyacceptable carrier. In a further aspect, a pharmaceutical compositioncan be provided comprising a therapeutically effective amount of atleast one disclosed compound. In a still further aspect, apharmaceutical composition can be provided comprising a prophylacticallyeffective amount of at least one disclosed compound. In yet a furtheraspect, the invention relates to pharmaceutical compositions comprisinga pharmaceutically acceptable carrier and a compound, wherein thecompound is present in an effective amount.

Pharmaceutically acceptable salts of the compounds are conventionalacid-addition salts or base-addition salts that retain the biologicaleffectiveness and properties of the compounds and are formed fromsuitable non-toxic organic or inorganic acids or organic or inorganicbases. Exemplary acid-addition salts include those derived frominorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodicacid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, andthose derived from organic acids such as p-toluenesulfonic acid,salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citricacid, malic acid, lactic acid, fumaric acid, and the like. Examplebase-addition salts include those derived from ammonium, potassium,sodium and, quaternary ammonium hydroxides, such as for example,tetramethylammonium hydroxide. Chemical modification of a pharmaceuticalcompound into a salt is a known technique to obtain improved physicaland chemical stability, hygroscopicity, flowability and solubility ofcompounds. See, e.g., H. Ansel et. al., Pharmaceutical Dosage Forms andDrug Delivery Systems (6th Ed. 1995) at pp. 196 and 1456-1457.

The pharmaceutical compositions comprise the compounds in apharmaceutically acceptable carrier. A pharmaceutically acceptablecarrier refers to sterile aqueous or nonaqueous solutions, dispersions,suspensions or emulsions, as well as sterile powders for reconstitutioninto sterile injectable solutions or dispersions just prior to use.Examples of suitable aqueous and nonaqueous carriers, diluents, solventsor vehicles include water, ethanol, polyols (such as glycerol, propyleneglycol, polyethylene glycol and the like), carboxymethylcellulose andsuitable mixtures thereof, vegetable oils (such as olive oil) andinjectable organic esters such as ethyl oleate. The compounds can beformulated with pharmaceutically acceptable carriers or diluents as wellas any other known adjuvants and excipients in accordance withconventional techniques such as those disclosed in Remington: TheScience and Practice of Pharmacy, 19th Edition, Gennaro, Ed., MackPublishing Co., Easton, Pa., 1995.

In a further aspect, the pharmaceutical composition is administered to amammal. In a still further aspect, the mammal is a human. In an evenfurther aspect, the human is a patient.

In a further aspect, the pharmaceutical composition is administeredfollowing identification of the mammal in need of treatment of cancer.In a still further aspect, the mammal has been diagnosed with a need fortreatment of cancer prior to the administering step.

In a further aspect, the pharmaceutical composition is administeredfollowing identification of the mammal in need of treatment of afibrotic disorder. In a still further aspect, the mammal has beendiagnosed with a need for treatment of a fibrotic disorder prior to theadministering step.

In a further aspect, the pharmaceutical composition is administeredfollowing identification of the mammal in need of immunotherapy. In astill further aspect, the mammal has been diagnosed with a need forimmunotherapy prior to the administering step.

In various aspects, the disclosed pharmaceutical compositions comprisethe disclosed compounds (including pharmaceutically acceptable salt(s)thereof) as an active ingredient, a pharmaceutically acceptable carrier,and, optionally, other therapeutic ingredients or adjuvants. The instantcompositions include those suitable for oral, rectal, topical, andparenteral (including subcutaneous, intramuscular, and intravenous)administration, although the most suitable route in any given case willdepend on the particular host, and nature and severity of the conditionsfor which the active ingredient is being administered. Thepharmaceutical compositions can be conveniently presented in unit dosageform and prepared by any of the methods well known in the art ofpharmacy.

The choice of carrier will be determined in part by the particularmethod used to administer the composition. Accordingly, there is a widevariety of suitable formulations of the pharmaceutical composition ofthe present invention. The following formulations for oral, aerosol,parenteral, subcutaneous, intravenous, intraarterial, intramuscular,intraperitoneal, intrathecal, rectal, and vaginal administration aremerely exemplary and are in no way limiting.

Formulations suitable for oral administration can consist of (a) liquidsolutions, such as an effective amount of the compound dissolved indiluents, such as water, saline, or orange juice; (b) capsules, sachets,tablets, lozenges, and troches, each containing a predetermined amountof the active ingredient, as solids or granule; (c) powders; (d)suspensions in an appropriate liquid; and (e) suitable emulsions. Liquidformulations may include diluents, such as water, cyclodextrin, dimethylsulfoxide and alcohols, for example, ethanol, benzyl alcohol, propyleneglycol, glycerin, and the polyethylene alcohols including polyethyleneglycol, either with or without the addition of a pharmaceuticallyacceptable surfactant, suspending agent, or emulsifying agent. Capsuleforms can be of the ordinary hard- or soft-shelled gelatin typecontaining, for example, surfactants, lubricants, and inert fillers,such as lactose, sucrose, calcium phosphate, and corn starch. Tabletforms can include one or more of the following: lactose, sucrose,mannitol, corn starch, potato starch, alginic acid, microcrystallinecellulose, acacia, gelatin, guar gum, colloidal silicon dioxide,croscarmellose sodium, talc, magnesium stearate, calcium stearate, zincstearate, stearic acid, and other excipients, colorants, diluents,buffering agents, disintegrating agents, moistening agents,preservatives, flavoring agents, and pharmacologically compatiblecarriers. Lozenge forms can comprise the active ingredient in a flavor,usually sucrose and acacia or tragacanth, as well as pastillescomprising the active ingredient in an inert base, such as gelatin andglycerin, or sucrose and acadia, emulsions, and gels containing, theaddition to the active ingredient in an inert base, such as gelatin andglycerin, or sucrose and acadia, emulsions, and gels containing, inaddition to the active ingredient, such carriers as are known in theart.

The compounds of the present disclosure alone or in combination withother suitable components, can be made into aerosol formulations to beadministered via inhalation. These aerosol formulations can be placedinto pressurized acceptable propellants, such asdichlorodifluoromethane, propane, and nitrogen. They also may beformulated as pharmaceuticals for non-pressured preparations, such as ina nebulizer or an atomizer.

Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and non-aqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.The compound can be administered in a physiologically acceptable diluentin a pharmaceutical carrier, such as a sterile liquid or mixture ofliquids, including water, saline, aqueous dextrose and related sugarsolutions, an alcohol, such as ethanol, isopropanol, or hexadecylalcohol, glycols, such as propylene glycol or polyethylene glycol suchas poly(ethyleneglycol) 400, glycerol ketals, such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, an oil, a fatty acid, a fatty acid esteror glyceride, or an acetylated fatty acid glyceride with or without theaddition of a pharmaceutically acceptable surfactant, such as a soap ora detergent, suspending agent, such as pectin, carbomers,methylcellulose, hydroxypropylmethylcellulose, orcarboxymethylcelluslose, or emulsifying agents and other pharmaceuticaladjuvants.

Oils which can be used in parenteral formulations include petroleum,animal, vegetable, or synthetic oils. Specific examples of oils includepeanut, soybean, sesame, cottonseed, corn, olive, petrolatum, andmineral. Suitable fatty acids for use in parenteral formulations includeoleic acid, stearic acid, and isostearic acid. Ethyl oleate andisopropyl myristate are examples of suitable fatty acid esters. Suitablesoaps for use in parenteral formulations include fatty alkali metal,ammonium, and triethanolamine salts, and suitable detergents include (a)cationic detergents such as, for example. dimethyldialkylammoniumhalides, and alkylpyridinium halides, (b) anionic detergents such as,for example, alkyl, aryl, and olefin sulfonates, alkyl olefin, ether,and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergentssuch as, for example, fatty amine oxides, fatty acid alkanolamides, andpolyoxyethylene polypropylene copolymers, (d) amphoteric detergents suchas, for example, alkyl β-aminopropionates, and 2-alkylimidazolinequaternary ammonium salts, and (e) mixtures thereof.

The parenteral formulations typically contain from about 0.5% to about25% by weight of the active ingredient in solution. Suitablepreservatives and buffers can be used in such formulations. In order tominimize or eliminate irritation at the site of injection, suchcompositions may contain one or more nonionic surfactants having ahydrophile-lipophile balance (HLB) of from about 12 to about 17. Thequantity of surfactant in such formulations ranges from about 5% toabout 15% by weight. Suitable surfactants include polyethylene sorbitanfatty acid esters, such as sorbitan monooleate and the high molecularweight adducts of ethylene oxide with a hydrophobic base, formed by thecondensation of propylene oxide with propylene glycol.

Pharmaceutically acceptable excipients are also well-known to those whoare skilled in the art. The choice of excipient will be determined inpart by the particular compound, as well as by the particular methodused to administer the composition. Accordingly, there is a wide varietyof suitable formulations of the pharmaceutical composition of thepresent disclosure. The following methods and excipients are merelyexemplary and are in no way limiting. The pharmaceutically acceptableexcipients preferably do not interfere with the action of the activeingredients and do not cause adverse side-effects. Suitable carriers andexcipients include solvents such as water, alcohol, and propyleneglycol, solid absorbants and diluents, surface active agents, suspendingagent, tableting binders, lubricants, flavors, and coloring agents.

The formulations can be presented in unit-dose or multi-dose sealedcontainers, such as ampules and vials, and can be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid excipient, for example, water, for injections,immediately prior to use. Extemporaneous injection solutions andsuspensions can be prepared from sterile powders, granules, and tablets.The requirements for effective pharmaceutical carriers for injectablecompositions are well known to those of ordinary skill in the art. SeePharmaceutics and Pharmacy Practice, J.B. Lippincott Co., Philadelphia,Pa., Banker and Chalmers, Eds., 238-250 (1982) and ASHP Handbook onInjectable Drugs, Toissel, 4^(th) ed., 622-630 (1986).

Formulations suitable for topical administration include lozengescomprising the active ingredient in a flavor, usually sucrose and acaciaor tragacanth; pastilles comprising the active ingredient in an inertbase, such as gelatin and glycerin, or sucrose and acacia; andmouthwashes comprising the active ingredient in a suitable liquidcarrier; as well as creams, emulsions, and gels containing, in additionto the active ingredient, such carriers as are known in the art.

Additionally, formulations suitable for rectal administration may bepresented as suppositories by mixing with a variety of bases such asemulsifying bases or water-soluble bases. Formulations suitable forvaginal administration may be presented as pessaries, tampons, creams,gels, pastes, foams, or spray formulas containing, in addition to theactive ingredient, such carriers as are known in the art to beappropriate.

One skilled in the art will appreciate that suitable methods ofexogenously administering a compound of the present disclosure to ananimal are available, and, although more than one route can be used toadminister a particular compound, a particular route can provide a moreimmediate and more effective reaction than another route.

As regards these applications, the present method includes theadministration to an animal, particularly a mammal, and moreparticularly a human, of a therapeutically effective amount of thecompound effective in the inhibition of TGF-β. The method also includesthe administration of a therapeutically effect amount of the compoundfor the treatment of patient having a predisposition for being afflictedwith a disorder associated with TGF-β activity. The dose administered toan animal, particularly a human, in the context of the present inventionshould be sufficient to affect a therapeutic response in the animal overa reasonable time frame. One skilled in the art will recognize thatdosage will depend upon a variety of factors including the condition ofthe animal, the body weight of the animal, as well as the severity andstage of the virus.

The total amount of the compound of the present disclosure administeredin a typical treatment is preferably between about 10 mg/kg and about1000 mg/kg of body weight for mice, and between about 100 mg/kg andabout 500 mg/kg of body weight, and more preferably between 200 mg/kgand about 400 mg/kg of body weight for humans per daily dose. This totalamount is typically, but not necessarily, administered as a series ofsmaller doses over a period of about one time per day to about threetimes per day for about 24 months, and preferably over a period of twiceper day for about 12 months.

The size of the dose also will be determined by the route, timing andfrequency of administration as well as the existence, nature and extentof any adverse side effects that might accompany the administration ofthe compound and the desired physiological effect. It will beappreciated by one of skill in the art that various conditions ordisease states, in particular chronic conditions or disease states, mayrequire prolonged treatment involving multiple administrations.

In a further aspect, the composition further comprises at least oneagent known to treat cancer. In a still further aspect, the cancer isselected from multiple myeloma and a hematologic malignancy.

In a further aspect, the composition further comprises at least oneagent known to have a side effect of increasing the risk of cancer.

In a further aspect, the composition further comprises at least oneagent known to treat a fibrotic disorder. In a still further aspect, thefibrotic disorder is found in the liver, the lung, the cardiac muscle,the kidney, the skin or the eye. In yet a further aspect, the fibroticdisorder is found in the liver. In an even further aspect, fibroticdisorder is glaucoma, amyotropic lateral sclerosis, or musculardystrophy.

In a further aspect, the composition further comprises at least oneagent known to have a side effect of increasing the risk of a fibroticdisorder.

In a further aspect, the composition further comprises at least oneagent known to treat an immune dysfunction.

In a further aspect, the composition further comprises at least oneagent known to have a side effect of increasing the risk of an immunedysfunction.

It is understood that the disclosed compositions can be prepared fromthe disclosed compounds. It is also understood that the disclosedcompositions can be employed in the disclosed methods of using.

D. Methods of Making the Compounds

In various aspects, the inventions relates to methods of makingcompounds useful to treat disorders associated with TGF-β activity suchas, for example, cancers, in particular, multiple myeloma andhematologic malignancies, immune dysfunction, and fibrotic disorders, inparticular, liver fibrosis, diabetic nephropathy, muscular dystrophy,amyotrophic lateral sclerosis, and glaucoma. Thus, in one aspect,disclosed are methods of making a disclosed compound.

Compounds according to the present disclosure can, for example, beprepared by the several methods outlined below. A practitioner skilledin the art will understand the appropriate use of protecting groups[see: Greene and Wuts, Protective Groups in Organic Synthesis] and thepreparation of known compounds found in the literature using thestandard methods of organic synthesis. There may come from time to timethe need to rearrange the order of the recommended synthetic steps,however this will be apparent to the judgment of a chemist skilled inthe art of organic synthesis. The following examples are provided sothat the invention might be more fully understood, are illustrativeonly, and should not be construed as limiting.

In one aspect, the disclosed compounds comprise the products of thesynthetic methods described herein. In a further aspect, the disclosedcompounds comprise a compound produced by a synthetic method describedherein. In a still further aspect, the invention comprises apharmaceutical composition comprising a therapeutically effective amountof the product of the disclosed methods and a pharmaceuticallyacceptable carrier. In a still further aspect, the invention comprises amethod for manufacturing a medicament comprising combining at least onecompound of any of disclosed compounds or at least one product of thedisclosed methods with a pharmaceutically acceptable carrier or diluent.

1. Route I

In one aspect, piperazine-2,5-diones can be prepared as shown below.

Compounds are represented in generic form, with substituents as noted incompound descriptions elsewhere herein, wherein PG is a protectinggroup, and wherein R is selected from methyl and ethyl. A more specificexample is set forth below.

In one aspect, compounds of type 1.10, and similar compounds, can beprepared according to reaction Scheme 1B above. Thus, compounds of type1.8 can be prepared by a coupling reaction of an appropriate carboxylicacid, e.g., 1.6 as shown above, and an appropriate amine, e.g., 1.7 asshown above. Appropriate carboxylic acids and appropriate amines arecommercially available or prepared by methods known to one skilled inthe art. The coupling reaction is carried out in the presence of anappropriate coupling agent, e.g.,1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU), and an appropriate base, e.g.,diisopropylethyl amine (DIPEA), in an appropriate solvent, e.g.,acetonitrile, for an appropriate period of time, e.g., 12 hours.Compounds of type 1.9 can be prepared by deprotection of an appropriateamine, e.g., 1.8 as shown above. The deprotection is carried out in thepresence of an appropriate acid, e.g., trifluoroacetic acid (TFA), in anappropriate solvent, e.g., dichloromethane, for an appropriate period oftime, e.g., 2 hours. Compounds of type 1.10 can be prepared bycyclization of an appropriate ester, e.g., 1.9 as shown above. Thecyclization is carried out in the presence of an appropriate acid, e.g.,acetic acid, in an appropriate solvent, e.g., n-butanol, at anappropriate temperature, e.g., 120-140° C., for an appropriate period oftime, e.g., 60-90 minutes hours, followed by addition of an appropriatebase, e.g., piperidine, in an appropriate solvent, e.g.,dimethylformamide, for an appropriate period of time, e.g., 2 hours. Ascan be appreciated by one skilled in the art, the above reactionprovides an example of a generalized approach wherein compounds similarin structure to the specific reactants above (compounds similar tocompounds of type 1.1, 1.2, 1.3, and 1.4), can be substituted in thereaction to provide substituted piperazine-2,5-diones similar to Formula1.5.

E. Methods of Using the Compounds

The compounds and pharmaceutical compositions of the invention areuseful in treating or controlling disorders associated with TGF-βactivity, in particular, cancers such as, for example, multiple myelomaand hematologic malignancies, immunotherapy, and fibrotic disorders suchas, for example, liver fibrosis, amyoptrophic lateral sclerosis,diabetic nephropathy, muscular dystrophy, and glaucoma.

Examples of cancers for which the compounds and compositions can beuseful in treating, include, but are not limited to carcinoma, lymphoma,blastoma, sarcoma, and leukemia. More particular examples of suchcancers include breast cancer, prostate cancer, colon cancer, squamouscell cancer, small-cell lung cancer, non-small cell lung cancer,gastrointestinal cancer, pancreatic cancer, cervical cancer, ovariancancer, peritoneal cancer, liver cancer, e.g., hepatic carcinoma,bladder cancer, colorectal cancer, endometrial carcinoma, kidney cancer,and thyroid cancer.

In various aspects, further examples of cancers are basal cellcarcinoma, biliary tract cancer; bone cancer; brain and CNS cancer;choriocarcinoma; connective tissue cancer; esophageal cancer; eyecancer; cancer of the head and neck; gastric cancer; intra-epithelialneoplasm; larynx cancer; lymphoma including Hodgkin's and Non-Hodgkin'slymphoma; melanoma; myeloma; neuroblastoma; oral cavity cancer (e.g.,lip, tongue, mouth, and pharynx); retinoblastoma; rhabdomyosarcoma;rectal cancer; cancer of the respiratory system; sarcoma; skin cancer;stomach cancer; testicular cancer; uterine cancer; cancer of the urinarysystem, as well as other carcinomas and sarcomas

Examples of fibrotic disorders for which the compounds and compositionscan be useful in treating, include, but are not limited to, pulmonaryfibrosis, glomerulonephritis, liver cirrhosis, diabetic nephropathy,proliferative vitreoretinopathy, systemic sclerosis, scleroderma,muscular dystrophy, amyotrophic lateral sclerosis, and glaucoma.

To treat or control the disorder, the compounds and pharmaceuticalcompositions comprising the compounds are administered to a subject inneed thereof, such as a vertebrate, e.g., a mammal, a fish, a bird, areptile, or an amphibian. The subject can be a human, non-human primate,horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent.The term does not denote a particular age or sex. Thus, adult andnewborn subjects, as well as fetuses, whether male or female, areintended to be covered. The subject is preferably a mammal, such as ahuman. Prior to administering the compounds or compositions, the subjectcan be diagnosed with a need for treatment of a cancer, immunedysfunction, or of a fibrotic disorder.

The compounds or compositions can be administered to the subjectaccording to any method. Such methods are well known to those skilled inthe art and include, but are not limited to, oral administration,transdermal administration, administration by inhalation, nasaladministration, topical administration, intravaginal administration,ophthalmic administration, intraaural administration, intracerebraladministration, rectal administration, sublingual administration, buccaladministration and parenteral administration, including injectable suchas intravenous administration, intra-arterial administration,intramuscular administration, and subcutaneous administration.Administration can be continuous or intermittent. A preparation can beadministered therapeutically; that is, administered to treat an existingdisease or condition. A preparation can also be administeredprophylactically; that is, administered for prevention of a cancer,immune dysfunction, or a fibrotic disorder.

The therapeutically effective amount or dosage of the compound can varywithin wide limits. Such a dosage is adjusted to the individualrequirements in each particular case including the specific compound(s)being administered, the route of administration, the condition beingtreated, as well as the patient being treated. In general, in the caseof oral or parenteral administration to adult humans weighingapproximately 70 Kg or more, a daily dosage of about 10 mg to about10,000 mg, preferably from about 200 mg to about 1,000 mg, should beappropriate, although the upper limit may be exceeded. The daily dosagecan be administered as a single dose or in divided doses, or forparenteral administration, as a continuous infusion. Single dosecompositions can contain such amounts or submultiples thereof of thecompound or composition to make up the daily dose. The dosage can beadjusted by the individual physician in the event of anycontraindications. Dosage can vary, and can be administered in one ormore dose administrations daily, for one or several days.

1. Treatment Methods

The compounds disclosed herein are useful for treating or controllingdisorders associated with TGF-β activity, in particular, cancers, immunedysfunction, and fibrotic disorders. Thus, provided is a methodcomprising administering a therapeutically effective amount of acomposition comprising a disclosed compound to a subject. In a furtheraspect, the method can be a method for treating cancer. In a stillfurther aspect, the method can be a method for treating a fibroticdisorder. In a still further aspect, the method can be a method fortreating immune dysfunction.

a. Treating Cancer

In one aspect, disclosed are methods of treating cancer associated withTGF-β activity in a mammal, the method comprising the step ofadministering to the mammal an effective amount of at least onedisclosed compound, or a pharmaceutically acceptable salt thereof.

In one aspect, disclosed are methods for treating cancer in a subject,the method comprising the step of administering to the subject aneffective amount of at least one compound having a structure representedby a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); or apharmaceutically acceptable salt thereof, thereby treating cancer in thesubject.

In one aspect, disclosed are methods for treating cancer in a subject,the method comprising the step of administering to the subject aneffective amount of at least one compound having a structure representedby a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b);provided that when n is 1 and each of R^(1a) and R^(1b) togethercomprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof, thereby treating cancer in thesubject.

In one aspect, disclosed are methods for treating cancer in a subject,the method comprising the step of administering to the subject aneffective amount of at least one compound having a structure selectedfrom:

or a pharmaceutically acceptable salt thereof, thereby treating cancerin the subject.

Examples of cancers include, but are not limited to, carcinoma,lymphoma, blastoma, sarcoma, and leukemia. More particular examples ofsuch cancers include breast cancer, prostate cancer, colon cancer,squamous cell cancer, small-cell lung cancer, non-small cell lungcancer, gastrointestinal cancer, pancreatic cancer, cervical cancer,ovarian cancer, peritoneal cancer, liver cancer, e.g., hepaticcarcinoma, bladder cancer, colorectal cancer, endometrial carcinoma,kidney cancer, and thyroid cancer.

In various aspects, further examples of cancers are basal cellcarcinoma, biliary tract cancer; bone cancer; brain and CNS cancer;choriocarcinoma; connective tissue cancer; esophageal cancer; eyecancer; cancer of the head and neck; gastric cancer; intra-epithelialneoplasm; larynx cancer; lymphoma including Hodgkin's and Non-Hodgkin'slymphoma; melanoma; myeloma; neuroblastoma; oral cavity cancer (e.g.,lip, tongue, mouth, and pharynx); retinoblastoma; rhabdomyosarcoma;rectal cancer; cancer of the respiratory system; sarcoma; skin cancer;stomach cancer; testicular cancer; uterine cancer; cancer of the urinarysystem, as well as other carcinomas and sarcomas

In a further aspect, the cancer is a hematological cancer. In a stillfurther aspect, the hematological cancer is selected from acute myeloidleukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloidleukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia,chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia(JMML), Hodgkin lymphoma, Non-Hodgkin lymphoma, multiple myeloma,solitary myeloma, localized myeloma, and extramedullary myeloma. In astill further aspect, the cancer is selected from chronic lymphocyticleukemia, small lymphocytic lymphoma, B-cell non-Hodgkin lymphoma, andlarge B-cell lymphoma.

In a further aspect, the cancer is a cancer of the brain. In a stillfurther aspect, the cancer of the brain is selected from a glioma,medulloblastoma, primitive neuroectodermal tumor (PNET), acousticneuroma, glioma, meningioma, pituitary adenoma, schwannoma, CNSlymphoma, primitive neuroectodermal tumor, craniopharyngioma, chordoma,medulloblastoma, cerebral neuroblastoma, central neurocytoma,pineocytoma, pineoblastoma, atypical teratoid rhabdoid tumor,chondrosarcoma, chondroma, choroid plexus carcinoma, choroid plexuspapilloma, craniopharyngioma, dysembryoplastic neuroepithelial tumor,gangliocytoma, germinoma, hemangioblastoma, hemangiopercytoma, andmetastatic brain tumor. In a yet further aspect, the glioma is selectedfrom ependymoma, astrocytoma, oligodendroglioma, and oligoastrocytoma.In an even further aspect, the glioma is selected fromjuvenile pilocyticastrocytoma, subependymal giant cell astrocytoma, ganglioglioma,subependymoma, pleomorphic xanthoastrocytom, anaplastic astrocytoma,glioblastoma multiforme, brain stem glioma, oligodendroglioma,ependymoma, oligoastrocytoma, cerebellar astrocytoma, desmoplasticinfantile astrocytoma, subependymal giant cell astrocytoma, diffuseastrocytoma, mixed glioma, optic glioma, gliomatosis cerebri, multifocalgliomatous tumor, multicentric glioblastoma multiforme tumor,paraganglioma, and ganglioglioma.

In one aspect, the cancer can be a cancer selected from cancers of theblood, brain, genitourinary tract, gastrointestinal tract, colon,rectum, breast, kidney, lymphatic system, stomach, lung, pancreas, andskin. In a further aspect, the cancer is selected from prostate cancer,glioblastoma multiforme, endometrial cancer, breast cancer, and coloncancer. In a further aspect, the cancer is selected from a cancer of thebreast, ovary, prostate, head, neck, and kidney. In a still furtheraspect, the cancer is selected from cancers of the blood, brain,genitourinary tract, gastrointestinal tract, colon, rectum, breast,liver, kidney, lymphatic system, stomach, lung, pancreas, and skin. In ayet further aspect, the cancer is selected from a cancer of the lung andliver. In an even further aspect, the cancer is selected from a cancerof the breast, ovary, testes, and prostate. In a still further aspect,the cancer is a cancer of the breast. In a yet further aspect, thecancer is a cancer of the ovary. In an even further aspect, the canceris a cancer of the prostate. In a still further aspect, the cancer is acancer of the testes.

In a further aspect, the cancer is selected from a cancer of the breast,cervix, gastrointestinal tract, colorectal tract, brain, skin, prostate,ovary, thyroid, testes, genitourinary tract, pancreas, and endometrias.In a still further aspect, the cancer is a cancer of the breast. In yeta further aspect, the cancer of the breast is a hormone resistantcancer. In an even further aspect, the cancer of the breast is a hormoneresistant cancer. In a still further aspect, the cancer is a cancer ofthe cervix. In yet a further aspect, the cancer is a cancer of theovary. In an even further aspect, the cancer is a cancer of theendometrias. In a still further aspect, the cancer is a cancer of thegenitourinary tract. In yet a further aspect, the cancer is a cancer ofthe colorectal tract. In an even further aspect, the cancer of thecolorectal tract is a colorectal carcinoma. In a still further aspect,the cancer is a cancer of the gastrointestinal tract. In yet a furtheraspect, the cancer of the gastrointestinal tract is a gastrointestinalstromal tumor. In an even further aspect, the cancer is a cancer of theskin. In a still further aspect, the cancer of the skin is a melanoma.In yet a further aspect, the cancer is a cancer of the brain. In an evenfurther aspect, the cancer of the brain is a glioma. In a still furtheraspect, the glioma is glioblastoma multiforme. In yet a further aspect,glioma is selected from is selected from an ependymoma, astrocytoma,oligodendroglioma, and oligoastrocytoma. In an even further aspect, thecancer of the brain is selected from acoustic neuroma, glioma,meningioma, pituitary adenoma, schwannoma, CNS lymphoma, primitiveneuroectodermal tumor, craniopharyngioma, chordoma, medulloblastoma,cerebral neuroblastoma, central neurocytoma, pineocytoma, pineoblastoma,atypical teratoid rhabdoid tumor, chondrosarcoma, chondroma, choroidplexus carcinoma, choroid plexus papilloma, craniopharyngioma,dysembryoplastic neuroepithelial tumor, gangliocytoma, germinoma,hemangioblastoma, and hemangiopercytoma. In a still further aspect, thehematological cancer is selected from a leukemia, lymphoma, chronicmyeloproliferative disorder, myelodysplastic syndrome,myeloproliferative neoplasm, and plasma cell neoplasm (myeloma). In yeta further aspect, the hematological cancer is leukemia. In an evenfurther aspect, the leukemia is selected from acute leukemia, acutelymphocytic leukemia, acute myelocytic leukemia, myeloblastic leukemia,promyelocytic leukemia, myelomonocytic leukemia, monocytic leukemia,erythroleukemia, chronic leukemia, chronic myelocytic (granulocytic)leukemia, and chronic lymphocytic leukemia. In a still further aspect,the leukemia is acute lymphocytic leukemia. In yet a further aspect, thehematological cancer is lymphoma. In an even further aspect, thehematological cancer is myeloma. In a still further aspect, the myelomais multiple myeloma.

In a further aspect, the carcinoma is selected from colon carcinoma,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, bile duct carcinoma,choriocarcinoma, seminoma, embryonal carcinoma, lung carcinoma, smallcell lung carcinoma, bladder carcinoma, and epithelial carcinoma.

In a further aspect, the cancer is selected from breast cancer, cervicalcancer, gastrointestinal cancer, colorectal cancer, brain cancer, skincancer, prostate cancer, ovarian cancer, thyroid cancer, testicularcancer, pancreatic cancer, endometrial cancer, melanoma, glioma,leukemia, lymphoma, chronic myeloproliferative disorder, myelodysplasticsyndrome, myeloproliferative neoplasm, and plasma cell neoplasm(myeloma).

In a further aspect, the subject has been diagnosed with a need fortreatment of cancer prior to the administering step.

In a further aspect, the subject is a mammal. In a still further aspect,the mammal is a human.

In a further aspect, the method further comprises the step ofidentifying a subject in need of treatment of cancer.

In a further aspect, the method further comprises the step ofadministering a therapeutically effective amount of at least onechemotherapeutic agent. Examples of chemotherapeutic agents include, butare not limited to, alkylating agents such as busulfan, cis-platin,mitomycin C, and carboplatin; antimitotic agents such as colchicine,vinblastine, paclitaxel (e.g., TAXOL®), and docetaxel; topoisomerase Iinhibitors such as camptothecin and topotecan; topoisomerase IIinhibitors such as doxorubicin and etoposide; RNA/DNA antimetabolitessuch as 5-azacytidine, 5-fluorouracil and methotrexate; DNAantimetabolites such as 5-fluoro-2′-deoxy-uridine, ara-C, hydroxyurea,gemcitabine, capecitabine and thioguanine; antibodies such as HERCEPTIN®and RITUXAN®, as well as other known chemotherapeutics such asphotofrin, melphalan, chlorambucil, cyclophosamide, ifosfamide,vincristine, mitoguazone, epirubicin, aclarubicin, bleomycin,mitoxantrone, elliptinium, fludarabine, octreotide, retinoic acid,tamoxifen and alanosine.

In a further aspect, the at least one compound and the at least oneagent are administered sequentially. In a still further aspect, the atleast one compound and the at least one agent are administeredsimultaneously.

In a further aspect, the at least one compound and the at least oneagent are co-formulated. In a still further aspect, the at least onecompound and the at least one agent are co-packaged.

b. Treating a Fibrotic Disorder

In one aspect, disclosed are methods of treating a fibrotic disorderassociated with TGF-β activity in a mammal, the method comprising thestep of administering to the mammal an effective amount of at least onedisclosed compound, or a pharmaceutically acceptable salt thereof.

In one aspect, disclosed are methods for treating a fibrotic disorder ina subject, the method comprising the step of administering to thesubject an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b);provided that when n is 1 and each of R^(1a) and R^(1b) togethercomprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof, thereby treating the fibroticdisorder in the subject.

In one aspect, disclosed are methods for treating a fibrotic disorder ina subject, the method comprising the step of administering to thesubject an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); or apharmaceutically acceptable salt thereof, thereby treating the fibroticdisorder in the subject.

In one aspect, disclosed are methods for treating a fibrotic disorder ina subject, the method comprising the step of administering to thesubject an effective amount of at least one compound having a structureselected from:

or a pharmaceutically acceptable salt thereof, thereby treating thefibrotic disorder in the subject.

Examples of fibrotic disorders for which the compounds and compositionscan be useful in treating, include, but are not limited to, pulmonaryfibrosis, diabetic nephropathy, glomerulonephritis, liver cirrhosis,muscular dystrophy, proliferative vitreoretinopathy, systemic sclerosis,scleroderma, amyotrophic lateral sclerosis, and glaucoma.

In a further aspect, the subject has been diagnosed with a need fortreatment of the disorder prior to the administering step.

In a further aspect, the subject is a mammal. In a still further aspect,the mammal is a human.

In a further aspect, the method further comprises the step ofidentifying a subject in need of treatment of the fibrotic disorder.

In a further aspect, the method further comprises the step ofadministering a therapeutically effective amount of at least one agentknown to treat a fibrotic disorder. In a still further aspect, the atleast one agent is selected from pirfenidone, nintedanib, aprostaglandin such as latanoprost and bimaotoprost, a beta blocker suchas timolol and betaxolol, an alpha-adrenergic agonist such asapraclonidine and brimonidine, a carbonic anhydrase inhibitor such asdorzolamide and brinzolamide, a moitic or cholinergic agent such aspilocarpine, a diuretic, an angiotenisin-converting enzyme (ACE)inhibitor, an angiotensin II receptor blocker, an anti-inflammatoryagent, and an anti-fibrotic agent.

In a further aspect, the at least one compound and the at least oneagent are administered sequentially. In a still further aspect, the atleast one compound and the at least one agent are administeredsimultaneously.

In a further aspect, the at least one compound and the at least oneagent are co-formulated. In a still further aspect, the at least onecompound and the at least one agent are co-packaged.

c. Treating an Immune Dysfunction

In one aspect, disclosed are methods of treating an immune dysfunctionassociated with TGF-β activity in a mammal, the method comprising thestep of administering to the mammal an effective amount of at least onedisclosed compound, or a pharmaceutically acceptable salt thereof.

In one aspect, disclosed are methods for treating an immune dysfunctionin a subject, the method comprising the step of administering to thesubject an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); or apharmaceutically acceptable salt thereof, thereby treating the immunedysfunction in the subject.

In one aspect, disclosed are methods for treating an immune dysfunctionin a subject, the method comprising the step of administering to thesubject an effective amount of at least one compound having a structureselected from:

or a pharmaceutically acceptable salt thereof, thereby treating theimmune dysfunction in the subject.

In a further aspect, the subject has been diagnosed with a need forimmunotherapy prior to the administering step. Examples of immunotherapyinclude, but are not limited to injection immunotherapy, topicalimmunotherapy, BCG immunotherapy, dendritic cell-based pump-priming,T-cell adoptive transfer, administration of an immunomodulator, immuneenhancement therapy, use of genetically engineered T-cells,antimicrobial immunotherapy, and immunosuppression.

In a further aspect, the subject is a mammal. In a still further aspect,the mammal is a human.

In a further aspect, the method further comprises the step ofidentifying a subject in need of immunotherapy.

In a further aspect, the method further comprises the step ofadministering a therapeutically effective amount of at least one agentknown to treat an immune dysfunction. Examples of agents known to treatimmune dysfunction include, but are not limited to, interleukins (i.e.,IL-2, IL-7, and IL-12), cytokines (i.e., interferons, G-CSF, imiquimod),chemokines (i.e., CCL3, CCL26, and CXCL7), immunomodulatory imide drugs(i.e., thalidomide, lenalidomide, pomalidomide, and apremilast),cytosine phosphate-guanosine, oligodeoxynulceotides, glucans, cytostaticdrugs, glucocorticoids, and immunosuppressive antibodies.

In a further aspect, the at least one compound and the at least oneagent are administered sequentially. In a still further aspect, the atleast one compound and the at least one agent are administeredsimultaneously.

In a further aspect, the at least one compound and the at least oneagent are co-formulated. In a still further aspect, the at least onecompound and the at least one agent are co-packaged.

2. Methods of Inhibiting TGF-B Activity in a Subject

In one aspect, disclosed are methods for inhibiting TGF-β activity in asubject, the method comprising the step of administering to the subjectan effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); orwherein R^(1a) is hydrogen, R^(1b) is —CH₂Cy³ and R⁴ is —C₆H₄R²¹;wherein Cy³ is selected from C3-C6 cycloalkyl, monocyclic heteroaryl,and aryl and is substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; or a pharmaceutically acceptable salt thereof, therebyinhibiting TGF-β activity in the subject.

In one aspect, disclosed are methods of inhibiting TGF-β activity in amammal, the method comprising the step of administering to the mammal atherapeutically effective amount of at least one disclosed compound, ora pharmaceutically acceptable salt thereof.

In one aspect, disclosed are methods for inhibiting TGF-β activity in asubject, the method comprising the step of administering to the subjectan effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); or apharmaceutically acceptable salt thereof, thereby inhibiting TGF-βactivity in the subject.

In one aspect, disclosed are methods for inhibiting TGF-β activity in asubject, the method comprising the step of administering to the subjectan effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b);provided that when n is 1 and each of R^(1a) and R^(1b) togethercomprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof, thereby inhibiting TGF-βactivity in the subject.

In one aspect, disclosed are methods for inhibiting TGF-β activity in asubject, the method comprising the step of administering to the subjectan effective amount of at least one compound having a structure selectedfrom:

or a pharmaceutically acceptable salt thereof, thereby inhibiting TGF-βactivity in the subject.

In a further aspect, the compound exhibits inhibition of TGF-β activity.In a still further aspect, the compound exhibits a decrease in TGF-βactivity.

In a further aspect, the subject is a mammal. In a still further aspect,the subject is a human.

In a further aspect, the subject has been diagnosed with a need fortreatment of the disorder prior to the administering step. In a stillfurther aspect, the method further comprises the step of identifying asubject in need of treatment of the disorder.

3. Methods of Inhibiting a TGF-B in at Least One Cell

In one aspect, disclosed are methods for inhibiting TGF-β activity in atleast one cell, the method comprising the step of contacting the atleast one cell with an effective amount of at least one disclosedcompound, or a pharmaceutically acceptable salt thereof.

In one aspect, disclosed are methods for inhibiting TGF-β activity in atleast one cell, the method comprising the step of contacting the cellwith an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), orwherein R^(1a) is hydrogen, R^(1b) is —CH₂Cy³ and R⁴ is —C₆H₄R²¹;wherein Cy³ is selected from C3-C6 cycloalkyl, monocyclic heteroaryl,and aryl and is substituted with 0-4 non-hydrogen groups independentlyselected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; or a pharmaceutically acceptable salt thereof, therebyTGF-β activity in the cell.

In one aspect, disclosed are methods for inhibiting TGF-β activity in atleast one cell, the method comprising the step of contacting the cellwith an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R^(31a), R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b), or apharmaceutically acceptable salt thereof, thereby TGF-β activity in thecell.

In one aspect, disclosed are methods for inhibiting TGF-β activity in atleast one cell, the method comprising the step of contacting the cellwith an effective amount of at least one compound having a structurerepresented by a formula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NHR²⁰ and Cy¹; wherein R²⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein Cy¹, when present, isselected from C5-C6 cycloalkyl, monocyclic heteroaryl, and —C₆H₄R²¹ andis substituted with 0-4 non-hydrogen groups independently selected fromhalogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4) dialkylamino; and whereinR²¹, when present, is selected from —(CH₂)_(q)NR^(31a)R^(31b) and—C(O)NR^(32a)R^(32b); wherein q, when present, is selected from 0 and 1;wherein each of R³, R^(31b), R^(32a), and R^(32b), when present, isindependently selected from hydrogen and C1-C4 alkyl; or wherein each ofR^(1a) and R^(1b) is hydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b),provided that when n is 1 and each of R^(1a) and R^(1b) togethercomprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is —NR^(20a)R^(20b)then each of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof, thereby TGF-β activity in thecell.

In one aspect, disclosed are methods for inhibiting TGF-β activity in atleast one cell, the method comprising the step of contacting the cellwith an effective amount of at least one compound selected from:

or a pharmaceutically acceptable salt thereof, thereby TGF-β activity inthe cell.

In a further aspect, the cell is mammalian. In a still further aspect,the cell is human. In yet a further aspect, the cell has been isolatedfrom a mammal prior to the contacting step.

In a further aspect, contacting is via administration to a mammal.

4. Use of Compounds

In one aspect, the invention relates to the use of a disclosed compoundor a product of a disclosed method. In a further aspect, a use relatesto the manufacture of a medicament for the treatment of cancer in amammal. In a still further aspect, a use relates to the manufacture of amedicament for the treatment of a fibrotic disorder in a mammal. In yeta further aspect, the use relates to the manufacture of a medicament forthe treatment of immune dysfunction.

Also provided are the uses of the disclosed compounds and products. Inone aspect, the invention relates to use of at least one disclosedcompound; or a pharmaceutically acceptable salt, hydrate, solvate, orpolymorph thereof. In a further aspect, the compound used is a productof a disclosed method of making.

In a further aspect, the use relates to a process for preparing apharmaceutical composition comprising a therapeutically effective amountof a disclosed compound or a product of a disclosed method of making, ora pharmaceutically acceptable salt, solvate, or polymorph thereof, foruse as a medicament.

In a further aspect, the use relates to a process for preparing apharmaceutical composition comprising a therapeutically effective amountof a disclosed compound or a product of a disclosed method of making, ora pharmaceutically acceptable salt, solvate, or polymorph thereof,wherein a pharmaceutically acceptable carrier is intimately mixed with atherapeutically effective amount of the compound or the product of adisclosed method of making.

In various aspects, the use relates to a treatment of a disorder in amammal. Also disclosed is the use of a compound for antagonism of TGF-βactivity. In one aspect, the use is characterized in that the mammal isa human. In one aspect, the use is characterized in that the disorder isa cancer or a fibrotic disorder. In one aspect, the use is characterizedin that the disorder relates to immune dysfunction.

In a further aspect, the use relates to the manufacture of a medicamentfor the treatment of cancer in a mammal. In a still further aspect, theuse relates to the manufacture of a medicament for the treatment of afibrotic disorder in a mammal. In a still further aspect, the userelates to the manufacture of a medicament for the treatment of immunedysfunction.

In a further aspect, the use relates to antagonism of a TGF-β activityin a mammal. In a further aspect, the use relates to modulating TGF-βactivity in a mammal. In a still further aspect, the use relates tomodulating TGF-β activity in a cell. In yet a further aspect, the mammalis a human.

It is understood that the disclosed uses can be employed in connectionwith the disclosed compounds, products of disclosed methods of making,methods, compositions, and kits. In a further aspect, the inventionrelates to the use of a disclosed compound or a disclosed product in themanufacture of a medicament for the treatment of cancer in a mammal. Ina further aspect, the cancer is selected from multiple myeloma andhematologic malignancy. In a further aspect, the invention relates tothe use of a disclosed compound or a disclosed product in themanufacture of a medicament for the treatment of a fibrotic disorder ina mammal. In a further aspect, the fibrotic disorder is liver fibrosis,diabetic nephropathy, muscular dystrophy, or glaucoma. In a stillfurther aspect, the use relates to the manufacture of a medicament forthe treatment of immune dysfunction.

5. Manufacture of a Medicament

In one aspect, the invention relates to a method for the manufacture ofa medicament for treating a disorder associated with TGF-β in a mammal,the method comprising combining a therapeutically effective amount of adisclosed compound or product of a disclosed method with apharmaceutically acceptable carrier or diluent.

As regards these applications, the present method includes theadministration to an animal, particularly a mammal, and moreparticularly a human, of a therapeutically effective amount of thecompound effective in the inhibition of TGF-β activity. The doseadministered to an animal, particularly a human, in the context of thepresent invention should be sufficient to affect a therapeutic responsein the animal over a reasonable time frame. One skilled in the art willrecognize that dosage will depend upon a variety of factors includingthe condition of the animal and the body weight of the animal.

The total amount of the compound of the present disclosure administeredin a typical treatment is preferably between about 10 mg/kg and about1000 mg/kg of body weight for mice, and between about 100 mg/kg andabout 500 mg/kg of body weight, and more preferably between 200 mg/kgand about 400 mg/kg of body weight for humans per daily dose. This totalamount is typically, but not necessarily, administered as a series ofsmaller doses over a period of about one time per day to about threetimes per day for about 24 months, and preferably over a period of twiceper day for about 12 months.

The size of the dose also will be determined by the route, timing andfrequency of administration as well as the existence, nature and extentof any adverse side effects that might accompany the administration ofthe compound and the desired physiological effect. It will beappreciated by one of skill in the art that various conditions ordisease states, in particular chronic conditions or disease states, mayrequire prolonged treatment involving multiple administrations.

Thus, in one aspect, the invention relates to the manufacture of amedicament comprising combining a disclosed compound or a product of adisclosed method of making, or a pharmaceutically acceptable salt,solvate, or polymorph thereof, with a pharmaceutically acceptablecarrier or diluent.

6. Kits

In one aspect, disclosed are kits comprising at least one disclosedcompound and one or more of: (a) at least one agent known to increaseTGF-β activity; (b) at least one agent known to treat cancer; (c) atleast one agent known to treat a fibrotic disorder; (d) at least oneagent known to treat an immune dysfunction; (e) instructions fortreating a disorder associated with TGF-β dysfunction; (f) instructionsfor treating cancer; (g) instructions for treating a fibrotic disorder;and (h) instructions for treating an immune dysfunction.

Examples of cancers for which the compounds and compositions can beuseful in treating, include, but are not limited to carcinoma, lymphoma,blastoma, sarcoma, and leukemia. More particular examples of suchcancers include breast cancer, prostate cancer, colon cancer, squamouscell cancer, small-cell lung cancer, non-small cell lung cancer,gastrointestinal cancer, pancreatic cancer, cervical cancer, ovariancancer, peritoneal cancer, liver cancer, e.g., hepatic carcinoma,bladder cancer, colorectal cancer, endometrial carcinoma, kidney cancer,and thyroid cancer.

In various aspects, further examples of cancers are basal cellcarcinoma, biliary tract cancer; bone cancer; brain and CNS cancer;choriocarcinoma; connective tissue cancer; esophageal cancer; eyecancer; cancer of the head and neck; gastric cancer; intra-epithelialneoplasm; larynx cancer; lymphoma including Hodgkin's and Non-Hodgkin'slymphoma; melanoma; myeloma; neuroblastoma; oral cavity cancer (e.g.,lip, tongue, mouth, and pharynx); retinoblastoma; rhabdomyosarcoma;rectal cancer; cancer of the respiratory system; sarcoma; skin cancer;stomach cancer; testicular cancer; uterine cancer; cancer of the urinarysystem, as well as other carcinomas and sarcomas

Examples of fibrotic disorders for which the compounds and compositionscan be useful in treating, include, but are not limited to, pulmonaryfibrosis, diabetic nephropathy, glomerulonephritis, liver cirrhosis,proliferative vitreoretinopathy, systemic sclerosis, scleroderma,muscular dystrophy, and glaucoma.

Examples of agents known to treat cancer include, but are not limitedto, alkylating agents such as busulfan, cis-platin, mitomycin C, andcarboplatin; antimitotic agents such as colchicine, vinblastine,paclitaxel (e.g., TAXOL®), and docetaxel; topoisomerase I inhibitorssuch as camptothecin and topotecan; topoisomerase II inhibitors such asdoxorubicin and etoposide; RNA/DNA antimetabolites such as5-azacytidine, 5-fluorouracil and methotrexate; DNA antimetabolites suchas 5-fluoro-2′-deoxy-uridine, ara-C, hydroxyurea, gemcitabine,capecitabine and thioguanine; antibodies such as HERCEPTIN® andRITUXAN®, as well as other known chemotherapeutics such as photofrin,melphalan, chlorambucil, cyclophosamide, ifosfamide, vincristine,mitoguazone, epirubicin, aclarubicin, bleomycin, mitoxantrone,elliptinium, fludarabine, octreotide, retinoic acid, tamoxifen andalanosine. Thus, in various aspects, an agent known to treat cancer ismelphalan.

Examples of agents known to treat fibrotic disorders include, but arenot limited to, pirfenidone, nintedanib, a prostaglandin such aslatanoprost and bimaotoprost, a beta blocker such as timolol andbetaxolol, an alpha-adrenergic agonist such as apraclonidine andbrimonidine, a carbonic anhydrase inhibitor such as dorzolamide andbrinzolamide, a moitic or cholinergic agent such as pilocarpine, adiuretic, an angiotenisin-converting enzyme (ACE) inhibitor, anangiotensin II receptor blocker, an anti-inflammatory agent, and ananti-fibrotic agent.

Examples of agents known to treat immune dysfunction include, but arenot limited to, interleukins (i.e., IL-2, IL-7, and IL-12), cytokines(i.e., interferons, G-CSF, imiquimod), chemokines (i.e., CCL3, CCL26,and CXCL7), immunomodulatory imide drugs (i.e., thalidomide,lenalidomide, pomalidomide, and apremilast), cytosinephosphate-guanosine, oligodeoxynulceotides, glucans, cytostatic drugs,glucocorticoids, and immunosuppressive antibodies.

In a further aspect, the at least one compound and the at least oneagent are co-formulated. In a further aspect, the at least one compoundand the at least one agent are co-packaged.

The kits can also comprise compounds and/or products co-packaged,co-formulated, and/or co-delivered with other components. For example, adrug manufacturer, a drug reseller, a physician, a compounding shop, ora pharmacist can provide a kit comprising a disclosed compound and/orproduct and another component for delivery to a patient.

It is understood that the disclosed kits can be prepared from thedisclosed compounds, products, and pharmaceutical compositions. It isalso understood that the disclosed kits can be employed in connectionwith the disclosed methods of using.

The foregoing description illustrates and describes the disclosure.Additionally, the disclosure shows and describes only the preferredembodiments but, as mentioned above, it is to be understood that it iscapable to use in various other combinations, modifications, andenvironments and is capable of changes or modifications within the scopeof the invention concepts as expressed herein, commensurate with theabove teachings and/or the skill or knowledge of the relevant art. Theembodiments described herein above are further intended to explain bestmodes known by applicant and to enable others skilled in the art toutilize the disclosure in such, or other, embodiments and with thevarious modifications required by the particular applications or usesthereof. Accordingly, the description is not intended to limit theinvention to the form disclosed herein. Also, it is intended to theappended claims be construed to include alternative embodiments.

All publications and patent applications cited in this specification areherein incorporated by reference, and for any and all purposes, as ifeach individual publication or patent application were specifically andindividually indicated to be incorporated by reference. In the event ofan inconsistency between the present disclosure and any publications orpatent application incorporated herein by reference, the presentdisclosure controls.

F. Examples

The following preparations and examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representative.

1. General Experimental Methods

The reactions were performed under a dry argon atmosphere and reactiontemperatures were measured externally. Anhydrous solvents over molecularsieves were purchased from Aldrich and used as such in reactions.Purification of all compounds was carried out by utilizing a TeledyneIsco Combiflash® Rf automated chromatography machine. Universal RediSepsolid sample loading pre-packed cartridges were used to absorb crudeproduct and purified on silica RediSep Rf Gold Silica (20-40 μmspherical silica) columns using appropriate solvent gradients. Puresamples were dried overnight under high vacuum over P₂O₅ at 78° C.before analyses. The reactions were monitored by thin-layerchromatography (TLC) on pre-coated silica gel (60F₂₅₄) aluminium plates(0.25 mm) from E. Merck and visualized using UV light (254 nm). Puresamples were dried overnight under high vacuum over P₂O₅ at 78° C.before analyses. The HR-mass spectral data were obtained on an AgilentLC-MSTOF by electrospray ionization (ESI). ¹H NMR spectra were recordedat 400 MHz on Agilent/Varian MR-400 spectrometer in CDCl₃ or DMSO-d₆ assolvents. The chemical shifts (δ) are in ppm downfield from standardtetramethylsilane (TMS). Coupling constants (J) are reported in Hertz(Hz). Chemical shifts (δ) listed for multiplets were measured from theapproximate centers, and relative integrals of peak areas agreed withthose expected for the assigned structures. ESI-MS spectra were recordedon a BioTof-2 time-of-flight mass spectrometer.

2. General Synthesis of Piperazine-2,5-Diones

Step 1: To a stirred mixture/solution of carboxylic acid and aminoacid-1 (1.1 eq.) in Acetonitrile/DMF was added DIPEA (2.5 eq.) followedby HATU (1.2 eq.) and the reaction mixture was stirred at rt for 12 h.Solvents were removed in vacuo and obtained residue was purified by IscoTeledyne Combiflash Rf200 using DCM and methanol as elutic solvents orin some cases solid crashed out, was filtered washed with diethyl etherand dried to afford the desired amide.

Step 2: To a stirred solution of amide in methylene chloride was addedTrifluoroacetic acid (10 eq.) at 0° C. under inert atmosphere and thereaction mixture was stirred at rt for 2 h. Solvent was removed in vacuoand obtained residue was co-evaporated with methylene chloride twice anddried under vacuum to afford residue. This residue was dried under highvacuum and used in next step without further purification.

Step 3: To this residue in a microwave vial with a stir bar was addedn-BuOH (0.15-0.2 M) followed by acetic acid (10 eq.) and the reactionmixture stirred for 5 min at room temperature. To this reaction mixturewas added Toluene (0.075-0.1 M) and the reaction mixture was irradiatedat 120/140° C. for 90 min. Solid crashed out, filtered, obtained solidwas washed with diethyl ether and dried to afford the desired carbamate.Or in some cases solvents were removed in vacuo and the residue waspurified by Isco Teledyne Combiflash Rf200 using DCM and methanol aselutic solvents to afford the desired carbamate.

Step 4: To a stirred solution of carbamate in DMF (0.5 M) was addedpiperidine (3 eq.) at rt under inert atmosphere and the reaction mixturewas stirred at rt for 2 h. Solvents were removed in vacuo and obtainedsolid was washed with diethyl ether to afford the desiredpiperazine-2,5-dione.

A. Preparation of (S)-3-(4-Aminobutyl)Piperazine-2,5-Dione

Step 1: General procedure was followed with(S)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and ethyl 2-aminoacetate as aminoacid-1 to obtain (S)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)acetateas a yellow solid, Yield 63.3%. ¹H NMR (400 MHz, Chloroform-d) δ7.80-7.73 (m, 2H), 7.60 (d, J=7.5 Hz, 2H), 7.40 (dddd, J=8.2, 7.5, 1.6,0.8 Hz, 2H), 7.30 (dd, J=1.0 Hz, 2H), 7.26 (d, 1H), 6.57 (s, 1H),5.04-5.11 (s, 1H), 4.94 (s, 1H), 4.40 (s, 2H), 4.20 (q, J=7.2 Hz, 3H),4.09-3.93 (m, 2H), 3.21 (d, J=6.8 Hz, 2H), 1.92-1.83 (m, 1H), 1.63-1.69(m, 1H), 1.52-1.56 (m, 2H), 1.44 (s, 10H), 1.26 (t, J=7.1 Hz, 4H).

Step 2: General procedure was followed to (S)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)acetatein Yield 91%. ¹H NMR (400 MHz, DMSO-d₆) δ 8.77 (t, J=5.9 Hz, 1H), 8.11(s, 2H), 7.82 (d, J=7.5 Hz, 2H), 7.60 (d, J=7.6 Hz, 2H), 7.40-7.32 (m,2H), 7.28 (td, J=7.4, 1.2 Hz, 2H), 7.13 (d, J=5.5 Hz, 1H), 4.35 (d,J=7.2 Hz, 1H), 4.24 (d, J=6.8 Hz, 3H), 4.14 (s, 3H), 2.91 (d, J=6.8 Hz,2H), 1.66 (s, 2H), 1.34 (d, J=6.9 Hz, 2H), 1.27-1.3 (m, 1H), 1.21-1.26(m, 2H), 1.11 (t, J=7.1 Hz, 3H).

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl (4-(3,6-dioxopiperazin-2-yl)butyl)carbamateas yellow color solid, Yield 37%, ¹H NMR (400 MHz, Methanol-d₄) δ 7.79(d, J=7.5 Hz, 2H), 7.64 (d, J=7.6 Hz, 2H), 7.38 (t, J=7.4 Hz, 2H), 7.30(td, J=7.4, 1.2 Hz, 2H), 4.35 (d, J=6.9 Hz, 2H), 4.18 (q, J=6.9 Hz, 1H),4.01-3.80 (m, 3H), 3.12 (q, J=6.9, 6.5 Hz, 2H), 1.85 (dq, J=19.8, 7.0,6.6 Hz, 1H), 1.51 (q, J=6.9, 6.4 Hz, 2H), 1.42 (s, 1H), 1.31-1.17 (m,1H), 0.87-0.96 (m, 1H). MS m/z 408 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-3-(4-aminobutyl)piperazine-2,5-dione as a yellow color solid, Yield88%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.16 (s, 1H), 7.96 (s, 1H), 3.78 (d,J=17.4 Hz, 1H), 3.73 (s, 1H), 3.66 (dd, J=17.0, 2.1 Hz, 1H), 2.94-2.86(m, 1H), 2.56 (t, J=6.5 Hz, 1H), 1.66 (p, J=6.2 Hz, 2H), 1.38-1.26 (m,4H).

b. Preparation of (S)-6-(4-Aminobutyl)-3,3-Dimethylpiperazine-2,5-Dione

Step 1: General procedure was followed with(S)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and ethyl 2-amino-2-methylpropanoate as amino acid-1 to obtain(S)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)-2-methylpropanoateas a off-white solid Yield-85%. ¹H NMR (400 MHz, DMSO-d₆) δ 8.09 (s,1H), 7.91-7.84 (m, 2H), 7.66 (d, J=7.5 Hz, 2H), 7.43-7.36 (m, 2H), 7.31(td, J=7.4, 1.1 Hz, 2H), 7.24 (t, J=5.7 Hz, 1H), 6.64 (d, J=8.5 Hz, 1H),4.27 (d, J=6.9 Hz, 2H), 4.18 (t, J=6.9 Hz, 1H), 3.96 (q, J=7.0 Hz, 2H),3.86 (d, J=7.1 Hz, 1H), 2.93 (d, J=6.2 Hz, 2H), 1.50 (s, 2H), 1.38-1.33(m, 11H), 1.31 (s, 3H), 1.29 (s, 3H), 1.23 (d, J=10.3 Hz, 2H), 1.09 (t,J=7.1 Hz, 3H). MS m/z 582 [M+H]⁺.

Step 2: General procedure was followed to obtain (S)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)-2-methylpropanoate,TFA as a sticky solid, Yield-62.5%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.68 (s,1H), 8.03 (s, 2H), 7.88 (dt, J=7.6, 1.0 Hz, 2H), 7.66 (d, J=7.5 Hz, 2H),7.45-7.38 (m, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.22 (t, J=5.6 Hz, 1H),4.29 (d, J=6.8 Hz, 2H), 4.19 (t, J=6.7 Hz, 1H), 4.02 (qd, J=7.1, 2.0 Hz,2H), 3.69-3.63 (1, 1H), 3.13 (qd, J=7.3, 4.2 Hz, 1H), 2.95 (q, J=6.6 Hz,1H), 1.67 (q, J=7.3 Hz, 2H), 1.39 (s, 3H), 1.34 (s, 3H), 1.30 (d, J=7.4Hz, 2H), 1.22 (d, J=7.3 Hz, 2H), 1.13 (t, J=7.1 Hz, 3H). MS m/z 482[M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-(5,5-dimethyl-3,6-dioxopiperazin-2-yl)butyl)carbamate as a whitesolid, Yield-22%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.17 (s, 1H), 7.96 (s,1H), 7.89-7.85 (m, 2H), 7.66 (d, J=7.6 Hz, 2H), 7.43-7.36 (m, 2H), 7.31(td, J=7.4, 1.2 Hz, 2H), 7.25 (t, J=5.7 Hz, 1H), 4.29-4.24 (m, 2H), 4.19(t, J=6.8 Hz, 1H), 3.87 (t, J=4.8 Hz, 1H), 3.15 (d, J=4.9 Hz, 1H),3.00-2.87 (m, 1H), 1.66 (s, 2H), 1.41-1.31 (m, 2H), 1.28 (d, J=2.1 Hz,7H), 1.19-1.25 (m, 1H). MS m/z 436 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-6-(4-aminobutyl)-3,3-dimethylpiperazine-2,5-dione as a white solid,Yield-84%, ¹H NMR (400 MHz, Methanol-d₄) δ 4.07 (t, J=5.1 Hz, 1H), 2.80(t, J=7.3 Hz, 2H), 1.97-1.74 (m, 2H), 1.61 (q, J=7.7 Hz, 2H), 1.54-1.45(m, 1H), 1.44 (d, J=2.2 Hz, 6H), 1.43-1.34 (m, 1H). HRMS m/z calcd forC₁₀H₁₉N₃O₂ [M+H]⁺: 213.1477, found: 213.1473.

c. Preparation of (S)-3-(4-Aminobenzyl)Piperazine-2,5-Dione

Step 1: General procedure was followed with(S)-3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoicacid and ethyl 2-aminoacetate, HCl as amino acid-1 to obtain (S)-ethyl2-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanamido)acetateas a white solid, Yield-74.3%. 1H NMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H),8.32 (t, J=5.9 Hz, 1H), 7.89 (dt, J=1.0, 7.5 Hz, 2H), 7.73 (d, J=7.4 Hz,2H), 7.48-7.37 (m, 2H), 7.33 (td, J=1.2, 7.5 Hz, 3H), 7.13 (d, J=8.2 Hz,2H), 6.85 (d, J=8.7 Hz, 1H), 4.44 (d, J=6.7 Hz, 2H), 4.28 (t, J=6.6 Hz,1H), 4.07 (q, J=7.1 Hz, 3H), 3.83 (qd, J=5.9, 17.5 Hz, 2H), 2.90 (dd,J=4.1, 13.8 Hz, 1H), 2.68-2.56 (m, 1H), 1.28 (s, 9H), 1.17 (t, J=7.1 Hz,3H). MS m z 488 [M−100 (for Boc)+1], 610 [M+Na]⁺.

Step 2: General procedure was followed to obtain (S)-ethyl2-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-aminopropanamido)acetate,AcO— as a sticky solid, Yield-89%, ¹H NMR (400 MHz, DMSO-d₆) δ 9.69 (s,1H), 8.89 (t, J=5.9 Hz, 1H), 8.08 (s, 2H), 7.89 (d, J=7.5 Hz, 2H), 7.72(d, J=7.4 Hz, 2H), 7.45-7.28 (m, 6H), 7.15 (d, J=8.1 Hz, 2H), 4.47 (d,J=6.7 Hz, 2H), 4.29 (t, J=6.5 Hz, 1H), 4.10 (q, J=7.1 Hz, 2H), 3.99 (d,J=7.8 Hz, 1H), 3.92 (dd, J=5.8, 3.8 Hz, 2H), 3.04 (dd, J=14.3, 5.2 Hz,1H), 2.86 (dd, J=14.3, 7.8 Hz, 1H), 1.18 (t, J=7.1 Hz, 3H). MS m/z 488[M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-((3,6-dioxopiperazin-2-yl)methyl)phenyl)carbamateas a white solid, Yield-63.3%, ¹H NMR (400 MHz, DMSO-d₆) δ 9.66 (s, 1H),8.10 (d, J=2.7 Hz, 1H), 7.93-7.83 (m, 3H), 7.73 (d, J=7.5 Hz, 2H),7.46-7.38 (m, 2H), 7.33 (td, J=7.5, 1.2 Hz, 4H), 7.03 (d, J=8.2 Hz, 2H),4.45 (d, J=6.7 Hz, 2H), 4.29 (t, J=6.6 Hz, 1H), 4.02-3.96 (m, 1H),3.37-3.33 (m, 1H), 3.01 (dd, J=13.6, 4.4 Hz, 1H), 2.84-2.72 (m, 2H). MSm/z 442 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-3-(4-aminobenzyl)piperazine-2,5-dione as a light yellow solid,Yield-72.6%, 1H NMR (400 MHz, Methanol-d4) δ 6.99-6.86 (m, 2H),6.69-6.61 (m, 2H), 4.16-4.05 (m, 1H), 3.39 (dd, J=17.6, 0.7 Hz, 1H),3.11 (dd, J=13.9, 3.8 Hz, 1H), 2.89-2.73 (m, 1H), 2.65 (dd, J=17.6, 1.1Hz, 1H). HRMS m/z calcd for C₁₁H₁₃N₃O₂[M+H]⁺: 219.1008, found: 219.1004.

d. Preparation of (S)-6-(4-Aminobenzyl)-3,3-Dimethylpiperazine-2,5-Dione

Step 1: General procedure was followed with(S)-3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoicacid and methyl 2-amino-2-methylpropanoate, HCl as amino acid to obtain(S)-ethyl2-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanamido)-2-methylpropanoateas a white fluffy solid, Yield-82%, 1H NMR (400 MHz, DMSO-d6) δ 9.60 (s,1H), 8.19 (s, 1H), 7.89 (dt, J=7.5, 1.0 Hz, 2H), 7.73 (d, J=7.4 Hz, 2H),7.41 (tdd, J=7.5, 1.2, 0.6 Hz, 2H), 7.33 (td, J=7.4, 1.2 Hz, 4H), 7.13(d, J=8.0 Hz, 2H), 6.72 (d, J=8.9 Hz, 1H), 4.44 (d, J=6.7 Hz, 2H), 4.28(t, J=6.7 Hz, 1H), 4.12-4.02 (m, 1H), 3.98 (q, J=7.0 Hz, 2H), 3.15 (d,J=5.2 Hz, 1H), 2.81 (dd, J=13.8, 4.6 Hz, 1H), 2.62 (dd, J=13.9, 10.2 Hz,1H), 1.31 (d, J=12.3 Hz, 6H), 1.28 (s, 8H), 1.11 (t, J=7.1 Hz, 3H). MSm/z 516 [(M−100) for Boc+H]⁺.

Step 2: General procedure was followed to obtain (S)-ethyl2-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-aminopropanamido)-2-methylpropanoate,TFA as a yield over two steps-49%; 1H NMR (400 MHz, DMSO-d6) δ 9.69 (s,1H), 8.67 (s, 1H), 8.06 (d, J=5.4 Hz, 2H), 7.90 (dt, J=7.7, 1.0 Hz, 2H),7.74-7.68 (m, 2H), 7.44-7.38 (m, 4H), 7.33 (td, J=7.4, 1.2 Hz, 2H), 7.16(d, J=8.3 Hz, 2H), 4.47 (d, J=6.6 Hz, 2H), 4.29 (t, J=6.6 Hz, 1H), 4.04(q, J=7.1 Hz, 2H), 3.90 (d, J=6.7 Hz, 1H), 3.01 (dd, J=14.1, 5.7 Hz,1H), 2.84 (dd, J=14.2, 8.0 Hz, 1H), 1.36 (s, 3H), 1.30 (s, 3H), 1.15 (t,J=7.1 Hz, 3H).

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-((5,5-dimethyl-3,6-dioxopiperazin-2-yl)methyl)phenyl)carbamate.1H NMR (400 MHz, DMSO-d6) δ 9.60 (s, 1H), 8.07 (s, 1H), 7.99 (d, J=1.9Hz, 1H), 7.89 (dt, J=7.5, 1.0 Hz, 2H), 7.72 (d, J=7.4 Hz, 2H), 7.44-7.37(m, 2H), 7.33 (td, J=7.5, 1.2 Hz, 4H), 7.03 (d, J=8.3 Hz, 2H), 4.44 (d,J=6.7 Hz, 2H), 4.28 (t, J=6.7 Hz, 1H), 4.14 (s, 1H), 3.06 (dd, J=13.6,3.6 Hz, 1H), 2.77 (dd, J=13.6, 4.9 Hz, 1H), 1.16 (s, 3H), 0.53 (s, 3H).MS m/z 470 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-6-(4-aminobenzyl)-3,3-dimethylpiperazine-2,5-dione as a white solid,Yield-62%, 1H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.89-7.84 (m, 1H),6.78 (d, J=8.0 Hz, 2H), 6.42 (d, J=8.0 Hz, 2H), 4.84 (s, 2H), 4.04 (s,1H), 2.94 (dd, J=13.7, 3.5 Hz, 1H), 2.64 (dd, J=13.5, 4.8 Hz, 1H), 1.15(s, 3H), 0.56 (s, 3H). HRMS m/z calcd for C₁₃H₁₇N₃O₂ [M+H]⁺: 247.1321,found: 247.1327.

e. Preparation of(S)-6-(3-(Aminomethyl)Benzyl)-3,3-Dimethylpiperazine-2,5-Dione

Step 1: General procedure was followed with(S)-3-(3-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanoicacid and methyl 2-amino-2-methylpropanoate, HCl to obtain(S)-ethyl2-(3-(3-(((((9H-fluoren-9-1)methoxy)carbonyl)amino)methyl)phenyl)-2-((tert-butoxycarbonyl)amino)propanamido)-2-methylpropanoateas a white solid, Yield-68%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.20 (s, 1H),7.87 (dd, J=7.5, 1.2 Hz, 2H), 7.83-7.74 (m, 1H), 7.68 (d, J=7.4 Hz, 2H),7.40 (td, J=7.4, 1.1 Hz, 2H), 7.37-7.26 (m, 2H), 7.18 (d, J=7.7 Hz, 1H),7.13 (s, 2H), 7.03 (d, J=7.6 Hz, 1H), 6.77 (d, J=8.8 Hz, 1H), 4.31 (d,J=7.5 Hz, 2H), 4.21 (t, J=6.9 Hz, 1H), 4.14 (d, J=5.9 Hz, 3H), 3.98 (dt,J=14.2, 6.6 Hz, 3H), 2.95-2.79 (m, 1H), 1.31 (s, 3H), 1.28 (s, 9H), 1.23(s, 3H), 1.10 (dd, J=8.0, 6.3 Hz, 3H). MS m/z 630 [M+H]⁺, 530 [(M−100)for Boc+H]⁺.

Step 2: General procedure was followed to obtain (S)-ethyl2-(3-(3-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)phenyl)-2-aminopropanamido)-2-methylpropanoateas a sticky solid, ¹H NMR (400 MHz, DMSO-d₆) δ 8.78 (s, 1H), 8.21 (s,2H), 7.98 (d, J=7.7 Hz, 2H), 7.88 (t, J=6.1 Hz, 1H), 7.78 (d, J=7.6 Hz,2H), 7.51 (t, J=7.5 Hz, 2H), 7.46-7.34 (m, 3H), 7.29-7.19 (m, 3H), 4.44(d, J=7.2 Hz, 2H), 4.30 (dd, J=23.8, 6.4 Hz, 3H), 4.13 (q, J=7.0 Hz,2H), 4.05 (s, 1H), 3.14 (dd, J=14.1, 5.9 Hz, 1H), 3.00 (dd, J=14.0, 7.7Hz, 1H), 1.45 (s, 3H), 1.38 (s, 3H), 1.29-1.20 (m, 3H). MS m/z 530[M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl3-((5,5-dimethyl-3,6-dioxopiperazin-2-yl)methyl)benzylcarbamate as awhite solid, Yield-54.8%, 1H NMR (400 MHz, DMSO-d6) δ 8.07 (s, 1H), 7.97(s, 1H), 7.86 (d, J=7.5 Hz, 2H), 7.75 (t, J=6.1 Hz, 1H), 7.68 (d, J=7.4Hz, 2H), 7.46-7.33 (m, 2H), 7.33-7.22 (m, 2H), 7.22-7.11 (m, 1H),7.10-6.97 (m, 3H), 4.30 (d, J=6.9 Hz, 2H), 4.26-4.16 (m, 2H), 4.10 (d,J=6.1 Hz, 2H), 3.08 (dd, J=13.5, 3.9 Hz, 1H), 2.85 (dd, J=13.5, 4.9 Hz,1H), 1.15 (s, 3H), 0.54 (s, 3H). MS m/z 484 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-6-(3-(aminomethyl)benzyl)-3,3-dimethylpiperazine-2,5-dione as awhite solid, Yield-76%, 1H NMR (400 MHz, DMSO-d6) δ 8.06 (s, 1H), 7.98(d, J=2.0 Hz, 1H), 7.17 (dd, J=5.9, 0.9 Hz, 2H), 7.09 (s, 1H), 7.06-6.94(m, 1H), 4.16 (ddd, J=4.9, 3.7, 1.9 Hz, 1H), 3.63 (s, 2H), 3.09 (dd,J=13.4, 3.8 Hz, 1H), 2.89-2.78 (m, 1H), 1.14 (s, 3H), 0.47 (s, 3H). HRMSm/z calcd for C₁₄H₁₉N₃O₂ [M+H]⁺: 261.1477, found: 261.1485.

f. Preparation of(S)-8-(4-Aminobutyl)-6,9-Diazaspiro[4.5]Decane-7,10-Dione

Step 1: General procedure was followed with(S)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and methyl 1-aminocyclopentanecarboxylate, HCl to obtain(S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)cyclopentanecarboxylateas a white solid, Yield-52%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.12 (s, 1H),7.86 (dt, J=7.7, 1.0 Hz, 2H), 7.65 (d, J=7.4 Hz, 2H), 7.45-7.35 (m, 2H),7.35-7.27 (m, 2H), 7.22 (t, J=5.7 Hz, 1H), 6.60 (d, J=8.4 Hz, 1H), 4.26(d, J=6.9 Hz, 2H), 4.17 (t, J=6.8 Hz, 1H), 3.85 (d, J=7.2 Hz, 1H), 3.50(s, 3H), 2.93 (d, J=7.0 Hz, 2H), 1.98 (dd, J=21.8, 12.3 Hz, 2H), 1.82(d, J=13.0 Hz, 2H), 1.60-1.62 (m, 4H), 1.44 (s, 2H), 1.37-1.30 (m, 11H),1.20-1.24 (m, 2H). MS m/z 594 [M+H]⁺.

Step 2: General procedure was followed to obtain (S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)cyclopentanecarboxylatein Yield-72% (Over step 2 and 3), ¹H NMR (400 MHz, DMSO-d₆) δ 8.74 (s,1H), 8.03 (s, 2H), 7.87 (dt, J=7.6, 1.0 Hz, 2H), 7.68-7.61 (m, 2H),7.42-7.36 (m, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.23 (t, J=5.7 Hz, 1H),4.28 (d, J=6.8 Hz, 2H), 4.18 (t, J=6.7 Hz, 1H), 3.54 (s, 3H), 2.95 (q,J=6.7 Hz, 2H), 1.93-1.79 (m, 2H), 1.66 (d, J=15.6 Hz, 6H), 1.43-1.36 (m,2H), 1.34-1.27 (m, 2H), 1.19-1.03 (m, 2H). MS m/z 494 [M+H]+

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-(7,10-dioxo-6,9-diazaspiro[4.5]decan-8-yl)butyl)carbamate as a whitesolid, 1H NMR (400 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.99 (d, J=2.0 Hz, 1H),7.87 (dt, J=7.6, 0.9 Hz, 2H), 7.66 (d, J=7.4 Hz, 2H), 7.40 (ddd, J=8.5,7.4, 1.0 Hz, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.24 (t, J=5.7 Hz, 1H),4.27 (d, J=6.9 Hz, 2H), 4.19 (t, J=6.9 Hz, 1H), 3.86-3.78 (m, 1H), 2.95(q, J=6.5 Hz, 2H), 2.04 (t, J=5.7 Hz, 2H), 1.64 (dt, J=14.1, 8.9 Hz,8H), 1.40-1.32 (m, 2H), 1.12 (s, 2H). MS m/z 462 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-8-(4-aminobutyl)-6,9-diazaspiro[4.5]decane-7,10-dione as a whitesolid, Yield-77%, 1H NMR (400 MHz, Deuterium Oxide) δ 4.06 (t, J=5.3 Hz,1H), 2.85 (t, J=7.7 Hz, 2H), 2.18-2.03 (m, 2H), 1.84-1.64 (m, 8H),1.64-1.48 (m, 2H), 1.41-1.18 (m, 2H). HRMS m/z calcd for C₁₂H₂₁N₃O₂[M+H]⁺: 239.1634, found: 239.1635.

g. Preparation of(S)-7-(4-Aminobutyl)-5,8-Diazaspiro[3.5]Nonane-6,9-Dione

Step 1: General procedure was followed with(S)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and methyl 1-aminocyclobutanecarboxylate to obtain (S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)cyclobutanecarboxylateas a white solid, Yield-55%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H),7.86 (dt, J=7.6, 1.0 Hz, 2H), 7.65 (d, J=7.4 Hz, 2H), 7.43-7.35 (m, 2H),7.35-7.27 (m, 2H), 7.23 (t, J=5.6 Hz, 1H), 6.63 (d, J=8.2 Hz, 1H), 4.26(d, J=6.9 Hz, 2H), 4.17 (t, J=6.9 Hz, 1H), 3.87 (q, J=7.5 Hz, 1H), 3.54(s, 3H), 2.94 (t, J=6.6 Hz, 2H), 2.66 (s, 2H), 2.39 (d, J=14.2 Hz, 1H),2.17-2.00 (m, 2H), 1.93-1.78 (m, 2H), 1.48 (dt, J=22.6, 7.6 Hz, 1H),1.34 (s, 10H), 1.23 (dd, J=6.6, 5.4 Hz, 3H). MS m/z 580 [M+H]⁺.

Step 2: General procedure was followed to obtain (S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)cyclobutanecarboxylatein Yield-68% (Over step 2 and 3), 1H NMR (400 MHz, DMSO-d6) δ 9.11 (s,1H), 8.04 (d, J=5.1 Hz, 2H), 7.87 (d, J=7.5 Hz, 2H), 7.65 (d, J=7.5 Hz,2H), 7.42-7.36 (m, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.22 (t, J=5.7 Hz,1H), 4.28 (d, J=6.8 Hz, 2H), 4.18 (t, J=6.8 Hz, 1H), 3.75-3.65 (m, 1H),3.58 (s, 3H), 2.95 (q, J=6.6 Hz, 2H), 2.67 (s, 2H), 2.21-2.03 (m, 2H),1.97-1.83 (m, 2H), 1.70 (q, J=7.4 Hz, 2H), 1.39 (q, J=7.1 Hz, 1H),1.32-1.18 (m, 3H). MS m z 480 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-(6,9-dioxo-5,8-diazaspiro[3.5]nonan-7-yl)butyl)carbamate as a whitesolid, Yield-77%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.63 (s, 1H), 8.00 (d,J=2.6 Hz, 1H), 7.87 (dt, J=7.6, 0.9 Hz, 2H), 7.66 (d, J=7.4 Hz, 2H),7.43-7.35 (m, 2H), 7.32 (dd, J=7.4, 1.2 Hz, 2H), 7.23 (t, J=5.7 Hz, 1H),4.26 (d, J=6.9 Hz, 2H), 4.18 (t, J=6.9 Hz, 1H), 3.69 (td, J=5.5, 2.5 Hz,1H), 2.94 (q, J=6.5 Hz, 2H), 2.5-2.52 (m, 1H), 2.44 (s, 1H), 2.13 (td,J=11.4, 10.7, 8.9 Hz, 2H), 1.91-1.65 (m, 2H), 1.57 (d, J=6.6 Hz, 2H),1.35 (q, J=7.0 Hz, 2H), 1.28-1.03 (m, 2H). MS m/z 470 [M+Na]⁺.

Step 4: General procedure was followed to obtain(S)-7-(4-aminobutyl)-5,8-diazaspiro[3.5]nonane-6,9-dione as a whitesolid, Yield-63.6%, ¹H NMR (400 MHz, Deuterium Oxide) δ 3.96 (t, J=5.4Hz, 1H), 2.73 (t, J=7.6 Hz, 2H), 2.65-2.43 (m, 2H), 2.30-2.11 (m, 2H),1.82 (pd, J=8.7, 4.3 Hz, 2H), 1.73-1.57 (m, 2H), 1.48 (p, J=7.6 Hz, 2H),1.36-1.09 (m, 2H). HRMS m/z calcd for C₁₁H₁₉N₃O₂ [M+H]⁺: 225.1480,found: 225.1477.

h. Preparation of(S)-7-(4-Aminobenzyl)-5,8-Diazaspiro[3.5]Nonane-6,9-Dione

Step 1: General procedure was followed with(S)-3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoicacid and methyl 1-aminocyclobutanecarboxylate to obtain (S)-methyl1-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanamido)cyclobutanecarboxylateas a off-white solid, Yield-55%, ¹H NMR (400 MHz, DMSO-d₆) δ 9.59 (s,1H), 8.63 (s, 1H), 8.13 (s, 1H), 7.89 (dt, J=7.7, 1.1 Hz, 2H), 7.72 (d,J=7.5 Hz, 2H), 7.40 (tdd, J=7.5, 1.2, 0.6 Hz, 2H), 7.32 (td, J=7.5, 1.2Hz, 3H), 7.11 (d, J=8.1 Hz, 2H), 6.71 (d, J=8.7 Hz, 1H), 4.44 (d, J=6.7Hz, 2H), 4.28 (t, J=6.7 Hz, 1H), 4.13-4.02 (m, 1H), 3.66-3.58 (m, 2H),3.56 (d, J=11.0 Hz, 3H), 3.18-3.06 (m, 2H), 2.82 (dd, J=13.8, 5.2 Hz,1H), 2.67-2.59 (m, 1H), 2.06 (q, J=9.0 Hz, 2H), 1.84 (s, 1H), 1.29 (s,9H). MS m/z 514 [(M−100) for Boc+H]⁺.

Steps 2 and 3: General procedure was followed to obtain (S)-methyl1-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-aminopropanamido)cyclobutanecarboxylate,TFA in Yield-63.6% (Over step 2 and 3), ¹H NMR (400 MHz, DMSO-d₆) δ 9.58(s, 1H), 8.53 (s, 1H), 7.95 (d, J=2.5 Hz, 1H), 7.88 (dt, J=7.6, 0.9 Hz,2H), 7.72 (d, J=7.4 Hz, 2H), 7.40 (tt, J=7.4, 1.0 Hz, 2H), 7.32 (td,J=7.4, 1.2 Hz, 4H), 6.98 (d, J=8.2 Hz, 2H), 4.43 (d, J=6.7 Hz, 2H), 4.27(t, J=6.6 Hz, 1H), 4.01 (td, J=4.6, 2.4 Hz, 1H), 2.95 (dd, J=13.6, 4.1Hz, 1H), 2.74 (dd, J=13.5, 4.9 Hz, 1H), 2.36 (ddd, J=12.4, 8.6, 4.3 Hz,1H), 1.98 (q, J=9.9, 8.6 Hz, 1H), 1.67-1.54 (m, 1H), 1.52-1.40 (m, 3H).MS m/z 482[M+H]⁺.

Step 4: General procedure was followed to obtain(S)-7-(4-aminobenzyl)-5,8-diazaspiro[3.5]nonane-6,9-dione as a whitesolid, Yield-46.4%, 1HNMR (400 MHz, Methanol-d₄) δ 6.94-6.86 (m, 2H),6.69-6.60 (m, 2H), 4.13 (t, J=4.2 Hz, 1H), 3.06 (dd, J=13.9, 3.9 Hz,1H), 2.78 (dd, J=13.9, 4.7 Hz, 1H), 2.57 (ddt, J=15.6, 8.9, 4.0 Hz, 1H),2.14-2.01 (m, 1H), 1.81-1.72 (m, 1H), 1.71-1.60 (m, 2H), 1.60-1.46 (m,1H). HRMS m/z calcd for C₁₄H₁₇N₃O₂ [M+H]⁺: 259.1321, found: 259.1322.

i. Preparation of(S)-8-(4-Aminobenzyl)-6,9-Diazaspiro[4.5]Decane-7,10-Dione

Step 1: General procedure was followed with(S)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and methyl 1-aminocyclopentanecarboxylate, HCl to obtain (S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)cyclopentanecarboxylateas an off-white solid, Yield-52%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.14 (s,1H), 7.87 (dt, J=7.6, 1.0 Hz, 2H), 7.66 (d, J=7.4 Hz, 2H), 7.47-7.36 (m,2H), 7.34-7.27 (m, 2H), 7.24 (t, J=5.7 Hz, 1H), 6.62 (d, J=8.3 Hz, 1H),4.26 (d, J=6.9 Hz, 2H), 4.18 (t, J=6.8 Hz, 1H), 3.89-3.82 (m, 1H), 3.50(s, 3H), 2.97-2.87 (m, 2H), 2.02-1.91 (m, 2H), 1.82 (dd, J=10.5, 4.2 Hz,2H), 1.61 (d, J=5.1 Hz, 5H), 1.50-1.37 (m, 3H), 1.30-1.33 (s, 11H). MSm/z 594 [M+H]⁺.

Step 2: General procedure was followed to obtain (S)-methyl1-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)cyclopentanecarboxylatein Yield-72% (Over step 2 and 3), ¹H NMR (400 MHz, DMSO-d₆) δ 8.74 (s,1H), 8.03 (s, 2H), 7.87 (dt, J=7.6, 1.0 Hz, 2H), 7.70-7.61 (m, 2H),7.46-7.35 (m, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.23 (t, J=5.7 Hz, 1H),4.28 (d, J=6.8 Hz, 2H), 4.18 (t, J=6.7 Hz, 2H), 3.54 (s, 3H), 2.95 (q,J=6.6 Hz, 2H), 1.93-1.77 (m, 2H), 1.64 (s, 6H), 1.44-1.27 (m, 4H),1.20-1.02 (m, 2H). MS m/z 494 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(4-(7,10-dioxo-6,9-diazaspiro[4.5]decan-8-yl)butyl)carbamate as a whitesolid, 1H NMR (400 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.99 (d, J=2.0 Hz, 1H),7.87 (dt, J=7.6, 0.9 Hz, 2H), 7.66 (d, J=7.4 Hz, 2H), 7.40 (ddd, J=8.5,7.4, 1.0 Hz, 2H), 7.31 (td, J=7.4, 1.2 Hz, 2H), 7.24 (t, J=5.7 Hz, 1H),4.27 (d, J=6.9 Hz, 2H), 4.19 (t, J=6.9 Hz, 1H), 3.86-3.78 (m, 1H), 2.95(q, J=6.5 Hz, 2H), 2.04 (t, J=5.7 Hz, 2H), 1.64 (dt, J=14.1, 8.9 Hz,8H), 1.40-1.32 (m, 2H), 1.12 (s, 2H). MS m/z 462 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-8-(4-aminobenzyl)-6,9-diazaspiro[4.5]decane-7,10-dione as aoff-white solid, Yield-98%, ¹H NMR (400 MHz, Methanol-d₄) δ 6.97-6.89(m, 2H), 6.72-6.62 (m, 2H), 4.20 (t, J=4.3 Hz, 1H), 3.11 (dd, J=13.9,4.0 Hz, 1H), 2.83 (dd, J=13.9, 4.6 Hz, 1H), 2.27-2.12 (m, 1H), 1.75-1.61(m, 2H), 1.61-1.50 (m, 3H), 1.42 (ddd, J=13.4, 8.6, 7.2 Hz, 1H), 0.80(dt, J=13.3, 6.5 Hz, 1H). HRMS m/z calcd for C₁₅H₁₉N₃O₂ [M+H]⁺:273.1477, found: 273.148.

j. Preparation of (R)-6-(4-Aminobutyl)-3,3-Dimethylpiperazine-2,5-Dione

Step 1: General procedure was followed with(R)-6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanoicacid and ethyl 2-amino-2-methylpropanoate, HCl to obtain (R)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)hexanamido)-2-methylpropanoateas a white solid, Yield-92%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.08 (s, 1H),7.90-7.83 (m, 2H), 7.66 (d, J=7.6 Hz, 2H), 7.39 (tdd, J=7.5, 1.2, 0.6Hz, 2H), 7.33-7.28 (m, 2H), 7.23 (t, J=5.7 Hz, 1H), 6.63 (d, J=8.4 Hz,1H), 4.26 (d, J=6.9 Hz, 2H), 4.18 (t, J=6.9 Hz, 1H), 3.96 (q, J=7.1 Hz,2H), 3.87 (s, 1H), 2.93 (s, 2H), 1.45 (s, 2H), 1.34 (s, 11H), 1.31 (s,3H), 1.29 (s, 3H), 1.24 (s, 2H), 1.09 (t, J=7.1 Hz, 3H). MS m/z 582[M+H]⁺.

Step 2: General procedure was followed to obtain (R)-ethyl2-(6-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminohexanamido)-2-methylpropanoate,TFA in Yield-49% (Over step 2 and 3)¹H NMR (400 MHz, Chloroform-d) δ7.76 (dt, J=7.6, 0.9 Hz, 2H), 7.54 (dq, J=7.5, 0.9 Hz, 3H), 7.44-7.36(m, 2H), 7.35-7.23 (m, 2H), 6.3 (s, 1H), 5.27 (s, 1H), 4.7-4.48 (bS,1H), 4.41 (s, 1H), 4.22-4.13 (m, 3H), 3.16 (s, 1H), 3.11-302 (m, 1H),1.87 (s, 2H), 1.56-1.53 (m, 1H), 1.5 (s, 3H), 1.48 (s, 3H), 1.41 (s,3H), 1.24 (t, J=7.1 Hz, 3H). MS m/z 482 [M+H]⁺.

Step 3: General procedure was followed to obtain(R)-(9H-fluoren-9-yl)methyl(4-(5,5-dimethyl-3,6-dioxopiperazin-2-yl)butyl)carbamate as a whitesolid, 1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.97 (s, 1H), 7.87 (dd,J=7.4, 1.0 Hz, 2H), 7.66 (d, J=7.4 Hz, 2H), 7.43-7.35 (m, 2H), 7.31 (td,J=7.5, 1.2 Hz, 2H), 7.26 (t, J=5.7 Hz, 1H), 4.27 (d, J=7.4 Hz, 2H), 4.18(t, J=6.9 Hz, 1H), 3.91-3.83 (m, 1H), 2.94 (q, J=6.5 Hz, 2H), 1.65 (tt,J=14.0, 7.3 Hz, 2H), 1.44-1.31 (m, 1H), 1.28 (d, J=2.2 Hz, 8H), 1.20 (d,J=20.7 Hz, 1H). MS m/z 436 [M+H]⁺.

Step 4: General procedure was followed to obtain(R)-6-(4-aminobutyl)-3,3-dimethylpiperazine-2,5-dione as a white solid,Yield-49%, ¹H NMR (400 MHz, Deuterium Oxide) δ 4.17-4.08 (m, 1H), 2.79(dd, J=8.5, 6.6 Hz, 2H), 1.87-1.77 (m, 1H), 1.76-1.66 (m, 1H), 1.53 (qd,J=8.1, 6.7 Hz, 2H), 1.37 (d, J=1.9 Hz, 7H), 1.31-1.18 (m, 1H). HRMS m/zcalcd for C₁₀H₁₉N₃O₂ [M+H]⁺: 213.1477, found: 213.1477.

k. Preparation of(S)-4-((5,5-Dimethyl-3,6-Dioxopiperazin-2-yl)Methyl)Benzamide

Step 1: General procedure was followed with(S)-2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanoic acidand methyl 2-amino-2-methylpropanoate, HCl to obtain (S)-methyl2-(2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanamido)-2-methylpropanoateas a white solid, Yield-73%, 1H NMR (400 MHz, Methanol-d4) δ 7.80 (d,J=7.9 Hz, 2H), 7.35 (d, J=7.9 Hz, 2H), 4.31 (dd, J=8.7, 6.0 Hz, 1H),3.66 (s, 3H), 3.18-3.01 (m, 1H), 2.86 (dd, J=13.9, 8.7 Hz, 1H), 1.43 (s,3H), 1.39 (s, 3H), 1.36 (s, 9H). MS m z 430 [M+Na]⁺.

Step 2: General procedure was followed to obtain (S)-methyl2-(2-amino-3-(4-carbamoylphenyl)propanamido)-2-methylpropanoate as asticky solid, 1H NMR (400 MHz, Methanol-d4) δ 7.92-7.84 (m, 2H),7.46-7.38 (m, 2H), 4.07 (dd, J=7.8, 6.7 Hz, 1H), 3.70 (s, 3H), 3.26 (dd,J=7.8, 6.2 Hz, 1H), 3.09 (dd, J=14.1, 7.8 Hz, 1H), 1.52-1.32 (m, 6H). MSm/z 308 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-4-((5,5-dimethyl-3,6-dioxopiperazin-2-yl)methyl)benzamide as a whitesolid, Yield-32.8%, 1H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 8.07 (d,J=1.9 Hz, 1H), 7.91 (s, 1H), 7.80-7.74 (m, 2H), 7.29 (s, 1H), 7.21 (d,J=8.3 Hz, 2H), 4.24 (ddd, J=5.3, 3.6, 1.8 Hz, 1H), 3.17 (dd, J=13.2, 3.7Hz, 1H), 2.89 (dd, J=13.3, 5.0 Hz, 1H), 1.16 (s, 3H), 0.51 (s, 3H). HRMSm/z calcd for C₁₄H₁₇N₃O₃ [M+H]⁺: 275.1270, found: 275.1272. HPLC-95.45%at t_(R)-1.87 min at 254 nm.

1. Preparation of (S)-4-((3,6-Dioxopiperazin-2-yl)Methyl)Benzamide

Step 1: General procedure was followed with(S)-2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanoic acidand ethyl 2-aminoacetate, HCl to obtain (S)-ethyl2-(2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanamido)acetateas a white solid, Yield-73%, 1H NMR (400 MHz, Methanol-d4) δ 7.90-7.69(m, 2H), 7.36 (d, J=8.3 Hz, 2H), 4.39 (dd, J=9.6, 5.0 Hz, 1H), 4.18 (q,J=7.1 Hz, 2H), 3.93 (d, J=6.9 Hz, 2H), 3.28-3.13 (m, 1H), 2.88 (dd,J=13.9, 9.6 Hz, 1H), 1.38-1.29 (m, 9H), 1.26 (t, J=7.1 Hz, 3H). MS m/z294 [(M−100) for Boc+H]⁺.

Step 2: General procedure was followed to obtain (S)-ethyl2-(2-amino-3-(4-carbamoylphenyl)propanamido)acetate as a sticky solid,1H NMR (400 MHz, DMSO-d6) δ 8.95 (t, J=5.8 Hz, 1H), 8.16 (d, J=5.6 Hz,2H), 8.09 (s, 1H), 7.95 (s, 1H), 7.87-7.79 (m, 2H), 7.37-7.30 (m, 2H),4.10 (q, J=7.1 Hz, 2H), 4.04-3.89 (m, 2H), 3.67 (d, J=5.6 Hz, 1H), 3.15(dd, J=14.1, 5.3 Hz, 1H), 2.99 (dd, J=14.1, 8.0 Hz, 1H), 1.33-1.06 (m,3H). MS m/z 294 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-4-((3,6-dioxopiperazin-2-yl)methyl)benzamide as a white solid,Yield-33.9%, 1H NMR (400 MHz, DMSO-d6) δ 8.18 (d, J=2.6 Hz, 1H), 7.92(s, 2H), 7.83-7.60 (m, 2H), 7.32 (s, 1H), 7.27-7.15 (m, 2H), 4.09 (d,J=2.8 Hz, 1H), 3.48-3.35 (m, 1H), 3.12 (dd, J=13.4, 4.6 Hz, 1H),2.99-2.79 (m, 2H). HRMS m/z calcd for C₁₂H₁₃N₃O₃ [M+H]⁺: 247.0957,found: 247.0961.

m. Preparation of(S)-4-((7,10-Dioxo-6,9-Diazaspiro[4.5]Decan-8-yl)Methyl)Benzamide

Step 1: General procedure was followed with(S)-2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanoic acidand methyl 1-aminocyclopentanecarboxylate to obtain (S)-methyl1-(2-((tert-butoxycarbonyl)amino)-3-(4-carbamoylphenyl)propanamido)cyclopentanecarboxylateas a white solid, Yield-68.6%, 1H NMR (400 MHz, Methanol-d4) δ 7.81 (d,J=8.0 Hz, 2H), 7.36 (d, J=8.1 Hz, 2H), 4.33 (dd, J=8.6, 6.3 Hz, 1H),3.64 (s, 3H), 3.09 (dd, J=13.7, 6.2 Hz, 1H), 2.88 (dd, J=13.8, 8.6 Hz,1H), 2.15 (s, 1H), 2.13 (s, 1H), 1.97-1.88 (m, 1H), 1.86 (dd, J=13.3,8.6 Hz, 1H), 1.73-1.56 (m, 4H), 1.37 (s, 9H). MS m/z 456 [M+Na]⁺.

Step 2: General procedure was followed to obtain (S)-methyl1-(2-amino-3-(4-carbamoylphenyl)propanamido)cyclopentanecarboxylate as asticky solid, 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.17 (d, J=5.4Hz, 2H), 7.95 (s, 1H), 7.84 (d, J=8.2 Hz, 2H), 7.37-7.29 (m, 2H), 4.01(q, J=6.3 Hz, 2H), 3.56 (s, 3H), 3.10 (dd, J=13.9, 6.2 Hz, 1H), 2.99(dd, J=14.0, 7.6 Hz, 1H), 2.12-1.94 (m, 2H), 1.84 (ddd, J=12.6, 7.4, 4.3Hz, 1H), 1.73 (ddd, J=12.4, 8.2, 4.2 Hz, 1H), 1.67-1.42 (m, 4H). MS m/z334 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-4-((7,10-dioxo-6,9-diazaspiro[4.5]decan-8-yl)methyl)benzamide as awhite solid, Yield-34.7%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.24 (s, 1H), 8.06(d, J=2.0 Hz, 1H), 7.91 (s, 1H), 7.79-7.72 (m, 2H), 7.28 (s, 1H),7.24-7.17 (m, 2H), 4.26-4.14 (m, 1H), 3.14 (dd, J=13.2, 4.2 Hz, 1H),2.93-2.84 (m, 1H), 2.03-1.91 (m, 1H), 1.66-1.55 (m, 1H), 1.53-1.45 (m,3H), 1.34 (ddt, J=20.7, 15.1, 6.4 Hz, 2H), 0.78-0.66 (m, 1H). HRMS m/zcalcd for C₁₆H₁₉N₃O₃ [M+H]⁺: 301.1426, found: 301.1422; HPLC-96.47% att_(R)-9.87 min at 240 nm.

n. Preparation of(S)-8-(3-Aminopropyl)-6,9-Diazaspiro[4.5]Decane-7,10-Dione

Step 1: General procedure was followed with(S)-5-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)pentanoicacid and methyl 1-aminocyclopentanecarboxylate to obtain (S)-methyl1-(5-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-((tert-butoxycarbonyl)amino)pentanamido)cyclopentanecarboxylateas a off-white solid, Yield-74.6%, ¹H NMR (400 MHz, Methanol-d₄) δ 7.79(d, J=7.5 Hz, 2H), 7.76-7.60 (m, 2H), 7.38 (tt, J=7.5, 1.0 Hz, 2H),7.34-7.28 (m, 2H), 4.34 (dd, J=6.9, 1.5 Hz, 2H), 4.20 (t, J=6.9 Hz, 1H),4.02 (d, J=7.9 Hz, 1H), 3.62 (s, 3H), 3.12 (hept, J=7.1 Hz, 2H),2.24-2.07 (m, 2H), 1.94 (dt, J=12.7, 6.2 Hz, 2H), 1.73 (p, J=4.1 Hz,5H), 1.61-1.45 (m, 3H), 1.43 (s, 9H). MS m/z 580 [M+H]⁺.

Step 2: General procedure was followed to obtain (S)-methyl1-(5-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2-aminopentanamido)cyclopentanecarboxylateas a fluffy solid, Yield-65% (over step 2 and 3), 1H NMR (400 MHz,Chloroform-d) δ 7.85 (s, 1H), 7.79-7.71 (m, 2H), 7.53 (d, J=7.5 Hz, 2H),7.39 (t, J=7.5 Hz, 2H), 7.29 (tdd, J=7.5, 2.1, 1.2 Hz, 2H), 4.39 (t,J=8.7 Hz, 1H), 4.30 (t, J=8.8 Hz, 1H), 4.23-4.08 (m, 2H), 3.63 (s, 3H),3.36-3.09 (m, 1H), 2.26-1.98 (m, 2H), 1.89 (s, 4H), 1.69 (s, 6H). MS m/z480 [M+H]⁺.

Step 3: General procedure was followed to obtain(S)-(9H-fluoren-9-yl)methyl(3-(7,10-dioxo-6,9-diazaspiro[4.5]decan-8-yl)propyl)carbamate, ¹H NMR(400 MHz, DMSO-d₆) δ 8.32 (s, 1H), 8.06-8.01 (m, 1H), 7.87 (d, J=7.5 Hz,2H), 7.66 (d, J=7.5 Hz, 2H), 7.46-7.36 (m, 2H), 7.31 (td, J=7.5, 2.5 Hz,3H), 4.26 (d, J=6.8 Hz, 2H), 4.19 (t, J=6.9 Hz, 1H), 3.87-3.80 (m, 1H),2.96 (q, J=6.7 Hz, 2H), 2.18-1.91 (m, 2H), 1.70 (d, J=7.5 Hz, 2H),1.68-1.55 (m, 6H), 1.45 (dq, J=15.5, 7.8, 7.1 Hz, 2H). MS m/z 448[M+H]⁺.

Step 4: General procedure was followed to obtain(S)-8-(3-aminopropyl)-6,9-diazaspiro[4.5]decane-7,10-dione as a whitesolid, Yield-63.6%, ¹H NMR (400 MHz, DMSO-d₆) δ 8.30 (d, J=5.5 Hz, 1H),8.09 (d, J=30.9 Hz, 2H), 6.67 (s, 1H), 3.82 (dd, J=7.0, 3.6 Hz, 1H),2.89 (q, J=6.7 Hz, 1H), 2.52 (m, 1H), 2.15-1.95 (m, 2H), 1.77-1.72 (m,1H), 1.72-1.69 (m, 1H), 1.68 (s, 1H), 1.66 (dd, J=4.2, 2.4 Hz, 1H), 1.64(d, J=3.7 Hz, 1H), 1.63-1.61 (m, 1H), 1.61-1.59 (m, 1H), 1.59-1.55 (m,1H), 1.38 (dt, J=15.8, 6.5 Hz, 2H). HRMS m/z calcd for C₁₁H₁₉N₃O₂[M+H]⁺: 225.1477, found: 225.1478.

o. Preparation of(S)-3-(4-Aminobenzyl)-1,4-Diazaspiro[5.5]Undecane-2,5-Dione

Step 1: General procedure was followed with(S)-3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoicacid and methyl 1-aminocyclohexanecarboxylate to obtain (S)-methyl1-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanamido)cyclohexanecarboxylateas a pale-yellow solid, Yield-86%, ¹H NMR (400 MHz, DMSO-d₆) δ 9.61 (s,1H), 7.96-7.89 (m, 2H), 7.89-7.84 (m, 2H), 7.73 (d, J=7.4 Hz, 2H), 7.41(tt, J=7.4, 0.9 Hz, 2H), 7.33 (td, J=7.5, 1.2 Hz, 3H), 7.14 (d, J=8.0Hz, 2H), 6.79 (d, J=8.8 Hz, 1H), 4.44 (d, J=6.7 Hz, 2H), 4.28 (t, J=6.6Hz, 1H), 4.15 (td, J=9.2, 4.7 Hz, 1H), 2.87 (s, 3H), 2.85-2.76 (m, 1H),2.65-2.60 (m, 1H), 1.91 (d, J=12.1 Hz, 2H), 1.62 (d, J=12.2 Hz, 2H),1.46 (s, 4H), 1.29 (s, 11H). MS m/z 664 [M+Na]⁺

Step 2: General procedure was followed to obtain (S)-methyl1-(3-(4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)phenyl)-2-aminopropanamido)cyclohexanecarboxylatein Yield-15% (over step 2 and 3), ¹H NMR (400 MHz, DMSO-d₆) δ 9.69 (s,1H), 8.44 (s, 1H), 8.07 (d, J=5.1 Hz, 2H), 7.95-7.86 (m, 3H), 7.72 (dd,J=7.5, 1.0 Hz, 2H), 7.41 (tt, J=7.4, 0.9 Hz, 3H), 7.33 (td, J=7.5, 1.2Hz, 2H), 7.18 (d, J=8.2 Hz, 2H), 4.47 (d, J=6.7 Hz, 2H), 4.29 (t, J=6.6Hz, 1H), 4.03 (d, J=10.3 Hz, 1H), 3.56 (s, 3H), 3.03 (dd, J=14.1, 5.9Hz, 1H), 2.86-2.79 (m, 1H), 1.91 (dd, J=14.4, 4.6 Hz, 1H), 1.83 (d,J=13.7 Hz, 1H), 1.75-1.58 (m, 2H), 1.43 (d, J=15.5 Hz, 4H), 1.23 (s,2H). MS m/z 542 [M+H]⁺.

Step 3:(S)-(9H-fluoren-9-yl)methyl(4-((2,5-dioxo-1,4-diazaspiro[5.5]undecan-3-yl)methyl)phenyl)carbamate,¹H NMR (400 MHz, DMSO-d₆) δ 9.59 (s, 1H), 8.05 (s, 1H), 7.96-7.83 (m,3H), 7.72 (d, J=7.5 Hz, 2H), 7.44-7.37 (m, 2H), 7.32 (td, J=7.4, 1.2 Hz,4H), 7.04 (d, J=8.2 Hz, 2H), 4.44 (d, J=6.7 Hz, 2H), 4.27 (t, J=6.7 Hz,1H), 4.14 (s, 1H), 3.05 (dd, J=13.5, 3.9 Hz, 1H), 2.79 (dd, J=13.7, 4.8Hz, 1H), 1.74-1.63 (m, 1H), 1.52 (dd, J=23.4, 13.2 Hz, 1H), 1.47-1.38(m, 2H), 1.35 (d, J=11.6 Hz, 2H), 1.29-1.18 (m, 2H), 1.10 (t, J=11.9 Hz,1H), 0.43 (d, J=12.5 Hz, 1H). MS m/z 510 [M+H]⁺.

Step 4: General procedure was followed to obtain(S)-3-(4-aminobenzyl)-1,4-diazaspiro[5.5]undecane-2,5-dione as a whitesolid, yield-60.8%, 1H NMR (400 MHz, Methanol-d4) δ 6.98-6.83 (m, 2H),6.70-6.54 (m, 2H), 4.21 (t, J=4.2 Hz, 1H), 3.13 (dd, J=13.9, 4.0 Hz,1H), 2.92-2.74 (m, 1H), 1.89 (td, J=14.1, 13.3, 4.2 Hz, 1H), 1.70-1.57(m, 1H), 1.57-1.46 (m, 3H), 1.46-1.35 (m, 1H), 1.35-1.13 (m, 3H),0.47-0.30 (m, 1H). HRMS m/z calcd for C₁₆H₂₁N₃O₂ [M+H]⁺: 287.1634,found: 287.1641.

3. Screening Assays for Compound Activity

For initial screening, an ELISA-based screening assay was used in whichpurified human platelet TSP-1 was incubated with compounds and thenincubated with recombinant latent TGF-β1 (purchased from R&D Systems).Activity is measured in a commercially available ELISA which detectsonly active TGF-β (R &D Systems). Activity in the presence of compoundsis compared to activity in TSP-1+latent TGF-β samples without inhibitorycompounds (Lu et al. (2016) Am J Pathol 186:678-690).

To specifically assess activity of compounds in liver cell specificassays, two different assays were used. In the first, human hepaticstellate cells were incubated overnight in low serum media to conditionthe media with secreted latent TGF-3. Purified TSP1 either preincubatedwith compounds or not will be added to cultures and then conditionedmedia harvested to assess TGF-β biological activity using the R&DSystems ELISA as above. Hepatic stellate cells will be purchased fromcommercial sources. Although hepatic stellate cells are the primarydrivers of liver fibrosis, conditions which drive liver fibrosis caninduce hepatocyte TSP-1 expression which can then have paracrine effectson controlling TGF-β activation in hepatic stellate cells. Furthermore,TGF-β can negatively impact hepatocyte regeneration following injury.Therefore, the ability of compounds to block TSP-1 activation of latentTGF-β will also be assessed in cultured primary human hepatocytesfollowing the protocol described for the hepatic stellate cells. Similarapproaches have successfully been used to screen compounds which inhibithuman myeloma cell derived latent TGF-β activation by TSP-1 (Lu et al.(2016) Am J Pathol 186:678-690).

4. TSP1 and TGF-B Decrease Osteoblast Differentiation and TSP1Inhibitory Peptide LSKL Increases Osteoblast Differentiation by MSCsUnder Osteogenic Conditions

Referring to FIG. 1A, MSCs were grown to confluence in basal (control)media. Cells were treated with control growth media, osteogenic media,or osteogenic media with TGF-β (5 ng/mL) or stripped TSP1 (10 nM),TSP1+LSKL (25 μM), TSP1+SLLK (25 μM control peptide), or TSP1+anti-TGF-β(5 μg/mL). Cultures were fed daily for 20 days. Alkaline phosphatasestaining is representative of triplicate wells.

Referring to FIG. 1B, cells were treated every 2 days for 20 days withMSC growth media (control) or osteogenic media with TGF-β (5 ng/mL) orTSP1 (10 nM)±daily treatment with 25 μM LSKL or SLLK peptides. RNAisolated from cells was used for RT-PCR analysis of Runx2 expression.Samples were run in duplicate and each experimental treatment intriplicate.

5. LSKL Peptide Treatment Reduces MM Tumor Burden in the SCID-Tibia MMModel

Referring to FIG. 2A and FIG. 2B, CAG human myeloma cells were injectedinto the intratibial marrow space of SCID mice. After 2 weeks, tumorswere imaged by bioluminescence and serum 1 g kappa levels measured. Micewere randomized to equalize 1 g kappa levels. Osmotic pumps wereimplanted subcutaneously to deliver saline or LSKL peptide (30mg/kg/day) (n=10/group). Tumors were imaged and serum 1 g kappa levelsmeasured at 2 and 4 weeks of treatment. Data are means±SEM.Bioluminescence; *p=0.019 ANOVA, serum kappa; p=0.037, t-test.

6. LSKL Reduces Phospho-Smad 2 in the Bone Marrow

Referring to FIG. 3A-C, at 4 weeks of treatment, tibiae from miceinjected with CAG MM cells were fixed and stained with antibody tophosphor-Smad 2. Left panels are from 2 different saline treated miceand the right panels are from LSKL-treated mice. Pixels of brownstaining were quantified in 4 fields per animal. 5-6 animals per groupwere analyzed. Data are the percent area exceeding the threshold forpositive staining (FIG. 3A). FIG. 3B represents the mean±SEM and FIG. 3Cshows the data for individuals. P=0.004.

7. TSP1 Induces TGF-B Activity in Cag MM Cells and LSKL Reduces TGF-BActivity

Referring to FIG. 4A, CAG MM cells were incubated with 30 nM TSP1 for 6hrs±LSKL or SLLK. Cell lysates were immunoblotted for phosphor-Smad 2.Blots were stripped and reprobed for total Smad 2/3 and GAPDH. Resultsare normalized to GAPDH (untreated controls=1). LSKL reduces TSP1induced treatment or luciferase activity in cells treated with activeTGF-β (not shown).

Referring to FIG. 4B, CAG-heparanase MM cells were treated with 67 nMTSP1 and conditioned media were assessed for TGF-β activity using PAI-1promoter luciferase reporter assay. There is ˜1.19 μM TGF-β in the addedTSP1 and LSKL blocks 80% of the CAG-heparanase MM cell generated TGF-βactivity.

8. Evaluation of Inhibitory Activity Against TGF-B

Table 1 below illustrates the effects of the disclosed compounds onTGF-β mediated by TSP-1.

TABLE 1 Inhibition of Inhibition of TGF-β mediated TGF-β mediated byTSP-1 in vitro by TSP-1 cell- Elisa assay at based assay at IC₅₀ No.Structure [Inhibitor] [Inhibitor] (nm)  1

At 10 pM = −67% At 50 pM = −4% At 100 pM = 37% At 10 nM = 38% At 100 nM= 59% At 1 μM = 74.5% ND  2

At 10 pM = 73% At 50 pM = 91% At 100 pM = 89% At 10 nM = 38% At 100 nM =71% At 1 μM = 76% ND  3

At 10 pM = 66% At 50 pM = 68% At 100 pM = 90% At 10 nM = 13% At 100 nM =32% At 1 μM = 75% ND  4

ND At 10 nM = 80% At 100 nM = 78% At 1 μM = 83% 1  5

ND At 10 nM = 45% At 100 nM = 85% At 1 μM = >100% 2.4  6

ND At 10 nM = 75% At 100 nM = 96% At 1 μM = 97% 1  7

ND At 10 nM = 72% At 100 nM = 89% At 1 μM = 92% 0.85  8

ND At 10 nM = 62% At 100 nM = 89% At 1 μM = 92% 4  9

ND At 10 nM = 40% At 100 nM = 92% At μM = >100% 11 10

ND Inactive ND 11

ND At 10 nM = 25% At 100 nM = 39% At 1 μM = 40% ND 12

ND At 10 nM = 40% At 100 nM = 65.6% At 1 μM = 68% ND 13

ND At 10 nM = 8% At 100 nM = 24.6% At 1 μM = 19.6% ND 14

ND At 10 nM = 54% At 100 nM = 85% At 1 μM = 90% ND 15

ND At 10 nM = 49% At 100 nM = 85% At 1 μM = 94% ND

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the present inventionwithout departing from the scope or spirit of the invention. Otheraspects of the invention will be apparent to those skilled in the artfrom consideration of the specification and practice of the inventiondisclosed herein. It is intended that the specification and examples beconsidered as exemplary only, with a true scope and spirit of theinvention being indicated by the following claims.

What is claimed is:
 1. A compound having a structure represented by aformula:

wherein n is selected from 1, 2, 3, and 4; wherein each of R^(1a) andR^(1b) is independently C1-C4 alkyl; or wherein each of R^(1a) andR^(1b) are optionally covalently bonded together and, together with theintermediate atoms, comprise a 3- to 7-membered cycloalkyl substitutedwith 0-4 non-hydrogen groups independently selected from halogen, —NH₂,C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4alkylamino, and (C1-C4)(C1-C4) dialkylamino; and wherein each of R² andR³ is independently selected from hydrogen and C1-C4 alkyl; and whereinR⁴ is selected from —NR^(20a)R^(20b) and Cy¹; wherein each of R^(20a)and R^(20b), when present, is independently selected from hydrogen,C1-C4 alkyl, C1-C4 haloalkyl, cycloalkyl, and heteroaryl; wherein Cy¹,when present, is selected from C5-C6 cycloalkyl, monocyclic heteroaryl,and —C₆H₄R²¹ and is substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4haloalkylh, C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and(C1-C4)(C1-C4) dialkylamino; and wherein R²¹, when present, is selectedfrom —(CH₂)_(q)NR^(31a)R^(31b) and —C(O)NR^(32a)R^(32b); wherein q, whenpresent, is selected from 0 and 1; wherein each of R^(31a), R^(31b),R^(32a), and R^(32b), when present, is independently selected fromhydrogen and C1-C4 alkyl; or wherein each of R^(1a) and R^(1b) ishydrogen and R⁴ is —C₆H₄C(O)NR^(32a)R^(32b); or wherein R^(1a) ishydrogen, R^(1b) is —CH₂Cy², and R⁴ is —C₆H₄R²¹; wherein Cy² is selectedfrom C3-C6 cycloalkyl substituted with 0-4 non-hydrogen groupsindependently selected from halogen, —NH₂, C1-C4 alkyl, C1-C4 haloalkyl,C1-C4 hydroxyalkyl, C1-C4 alkoxy, C1-C4 alkylamino, and (C1-C4)(C1-C4)dialkylamino; provided that when n is 1 and each of R^(1a) and R^(1b)together comprise a 3-membered cycloalkyl then Cy¹, when present, is notmonocyclic heteroaryl, and provided that when R⁴ is NR^(20a)R^(20b) theneach of R^(1a) and R^(1b) is independently C1-C4 alkyl, or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1,wherein n is
 1. 3. The compound of claim 1, wherein each of R² and R³ ishydrogen.
 4. The compound of claim 1, wherein R⁴ is NHR^(20a).
 5. Thecompound of claim 1, wherein each of R^(20a) and R^(20b) is hydrogen. 6.The compound of claim 1, wherein R⁴ is Cy¹.
 7. The compound of claim 1,having a structure represented by a formula:

wherein Z is selected from O, S, and NR⁴⁰; wherein R⁴⁰, when present, isselected from hydrogen and C1-C4 alkyl; wherein each of Q¹ and Q² isindependently selected from CH and N; and wherein R³³ is selected fromhydrogen and C1-C4 alkyl.
 8. The compound of claim 1, having a structurerepresented by a formula:

wherein R³³ is selected from hydrogen and C1-C4 alkyl.
 9. The compoundof claim 1, having a structure represented by a formula:

wherein Q¹ is selected from CH and N; and wherein R³³ is selected fromhydrogen and C1-C4 alkyl.
 10. The compound of claim 1, having astructure selected from:


11. The compound of claim 1, having a structure selected from:


12. A pharmaceutical composition comprising a therapeutically effectiveamount of at least one compound of claim 1 and a pharmaceuticallyacceptable carrier.